Latest revision as of 19:43, 17 October 2018

Transforming the ligation product of the Curcumin bio-sensor into E. coli BL21 DE3 Cultivating the culture, preparing E. coli culture glycerol stocks, and doing the mini preparation.
Sample

fresh colony of E. coli BL21 DE3 carrying the backbone pET30a containing αS1-casein + GS linker and T7 promotor gene from an overnight plate

July 6

None

July 7

Expression of αS1-casein

Cultivating E. coli culture containing target protein αS1-casein from the E. coli glycerol stock and inducing it with IPTG to produce αS1-casein.
Purifying the protein by His-Tag.

July 8

Growth curve exp

Observe the growth population of Bacillus subtilis in different pH Condition: pH = 4, 5, 6, 7, 8

July 9

None

July 10

Chip Production

Preparing 18 chips for bio-sensor sensitivity pretest
1. Dip the gold chips in 10mM Mua, RT for 4hrs.
2. Wash the chips with 95% EtOH three times and dry.
3. Add EDC+NHS mixture (100+100mM in DDW) on chips, RT for 1hrs.
4. DDW rinse the chips and dry.
5. Add αS1-casein on chips, RT for 1hrs.
6. Wash with PBS three times and dry.
6. Dip the chips in blocking solution, RT for 1.5hrs.
7. Wash with PBS three times and dry.

July 11

Bio-sensor sensitivity pretest

Using DPV(Differential Pulse Voltammetry) to check whether our bio-sensor can detect curcumin.
1. Add the diluted curcumin samples on our bio-sensor to react for 30min.
2. Rinse with wash buffer and dry the chips.
3. Wash the reference and counter electrodes with DDW, and dry them.
4. Set up the three electrodes system within electrocheFunctional analysisal cell.
5. Use the prototype of electrocheFunctional analysisal machine to measure the DPV method

July 12

Bio-sensor sensitivity assay –Determine Standard Curve and Create the Formula

Detecting the diluted standard curcumin samples by curcumin bio-sensor and made the standard curve and doing the polynomial curve fitting.

July 13

Bio-sensor sensitivity assay -Detecting the real Samples from Turmeric Root

Milling the turmeric root and dividing the powder into two groups. One of them was added with extraction buffer but not underwent the extraction protocol, and the other was added with extraction buffer but underwent the extraction process

July 14

None

July 15

Expression

Preparing competent cell E. coli ER2566

Growth curve exp

Observe the growth population of Bacillus subtilis in different salinity condition: 0.17M, 0.25M, 0.5M, 0.75M, 1.0M

July 16

Cloning

Amplify the insert gene (Pfu PCR)
Electrophoresis (To check PCR products)
Sample

Lacticin Z + pTXB1
Bovincin HJ50+ pTXB1

July 17

Cloning

Digestion of PCR products (Including insert and backbone pTXB1)
Ligation of digestion products (Ligase the DNA fragment into backbone pTXB1)
Sample

Lacticin Z + pTXB1
Bovincin HJ50+ pTXB1

July 18

Cloning

Transformation of ligation products (Transform into E. coli DH5α)
Taq PCR (PCR of ligation products to amplify insert gene)
Electrophoresis (To check PCR products)
Sample

Lacticin Z + pTXB1
Bovincin HJ50+ pTXB1

July 19

Cloning

Cultivation
Miniprep (Purify Plasmid)
Sample

Lacticin Z + pTXB1
Bovincin HJ50+ pTXB1

July 20

Cloning

Sequencing plasmid
Sample

Lacticin Z + pTXB1
Bovincin HJ50+ pTXB1

July 21

None

July 22

Cloning

Amplify the insert gene (Pfu PCR)
Electrophoresis (To check PCR products)
Digestion of PCR products (Including insert and backbone pTXB1)
Ligation of digestion products (Ligase the DNA fragment into backbone pTXB1)
Sample

Leucocyclicin Q + pTXB1

July 23

Cloning

Transformation of ligation products (Transform into E. coli DH5α)
Taq PCR (PCR of ligation products to amplify insert gene)
Electrophoresis (To check PCR products)
Sample

Leucocyclicin Q + pTXB1

July 24

Cloning

Cultivation
Miniprep (Purify Plasmid)
Sample

Leucocyclicin Q + pTXB1

July 25

Cloning

Sequencing plasmid
Sample

Leucocyclicin Q + pTXB1

July 26

None

July 27

None

July 28

None

July 29

Expression

Transformation (Transform correct plasmid into E. coli ER2566)

Leucocyclicin Q + pTXB1

July 30

None

July 31

None

August 1

None

August 2

None

August 3

None

August 4

None

August 5

Expression

Transformation (Transform correct plasmid into E. coli ER2566)
sample

Leucocyclicin Q + pTXB1

August 6

Expression

Cultivation (Cultivation of E. coli ER2566 colonies for IPTG induction)
IPTG Induction (To test the O.D. levels of E. coli ER2566 to induce)Condition: O.D. = 0.5, 1.3
sample

Leucocyclicin Q + pTXB1

August 7

Expression

SDS-PAGE (To check the protein production after induction)
sample

Leucocyclicin Q + pTXB1

August 8

None

August 9

None

August 10

None

August 11

Cloning

Amplify the insert gene (Pfu PCR)
Electrophoresis (To check PCR products)
sample

Enterocin B
Enterocin 96
Durancin TW49M

August 12

Cloning

Digestion of PCR products (Including insert and backbone pTXB1)
Ligation of digestion products (Ligase the DNA fragment into backbone pTXB1)
Transformation of ligation products (Transform into E. coli DH5α)
sample

Enterocin B + pTXB1
Enterocin 96 + pTXB1
Durancin TW-49M + pTXB1

Growth curve

Observe the growth of Bacillus subtilis in different temperature, pH, salinityCondition:
1.Temp: 37°C , pH:7 , salinity: 0.17M
2.Temp: 30°C , pH:9 , salinity: 0.5M
3.Temp: 25°C , pH:5 , salinity: 0.25M

August 13

Cloning

Taq PCR (PCR of ligation products to amplify insert gene)
Electrophoresis (To check PCR products)
sample

Enterocin B + pTXB1
Enterocin 96 + pTXB1
Durancin TW-49M + pTXB1

August 14

Cloning

Cultivation
Miniprep (Purify Plasmid)
Sample

Enterocin B + pTXB1
Enterocin 96 + pTXB1
Durancin TW-49M + pTXB1

Expression

Transformation (Transform correct plasmid into E. coli ER2566)to test different cultivation temperature

August 15

Cloning

Sequencing plasmid
Sample

Enterocin B + pTXB1
Enterocin 96 + pTXB1
Durancin TW-49M + pTXB1

Expression

Cultivation (Cultivation of E. coli ER2566 colonies for IPTG induction)
IPTG Induction (To test the temperature of cultivation after induction)Condition: temp = 37, 30, 13.5°C

August 16

Expression

SDS-PAGE (To check the protein production after induction)Check different temperature
IPTG Induction (To test the concentration of IPTG)Condition: concentration = 200, 400, 600, 800, 1000μM

August 17

Expression

SDS-PAGE (To check the protein production after induction) -> Check different IPTG concentration

August 18

Transformation

Transforming backbone pTXB1 into E. coli ER2566 to produce protein as our negative control in verifying the function of our target peptide.
Transforming backbone pTXB1 containing bacteriocin gene into E. coli ER2566 to produce our target protein as a bio-stimulator.
Sample

backbone pTXB1 mini
backbone pTXB1 containing Enterocin B gene mini
backbone pTXB1 containing Enterocin 96 gene mini
backbone pTXB1 containing Leucocyclicin Q gene mini

Cultivation

Cultivate the E. coli ER2566 at 37°C cultivating Bacillus subtilis ER2566 at 37°C carrying the backbone pTXB1 to produce protein as our negative control in verifying the function of our target peptide.
Cultivate the E. coli ER2566 at 37°C cultivating E. coli ER2566 at 37°C carrying the backbone pTXB1 containing bacteriocin gene to produce our target protein as a bio-stimulator.
Sample

Fresh colony of E. coli ER2566 carrying the backbone pTXB1 from an overnight plate
Fresh colony of E. coli ER2566 carrying the backbone pTXB1 containing Enterocin B gene from an overnight plate
Fresh colony of E. coli ER2566 carrying the backbone pTXB1 containing Enterocin 96 gene from an overnight plate
Fresh colony of E. coli ER2566 carrying the backbone pTXB1 containing Leucocyclicin Q gene from an overnight plate

IPTG Induction

Inducting E. coli ER2566 carrying the backbone pTXB1 after cultivating at 37°C to produce protein as our negative control in verifying the function of our target peptide
Inducting E. coli ER2566 carrying the backbone pTXB1 containing bacteriocin gene after cultivating at 37°C to produce our target protein as a bio-stimulator.
Sample

Culture of E. coli ER2566 carrying the backbone pTXB1
Culture of E. coli ER2566 carrying the backbone pTXB1 containing Enterocin B gene
Culture of E. coli ER2566 carrying the backbone pTXB1 containing Enterocin 96 gene
Culture of E. coli ER2566 carrying the backbone pTXB1 containing Leucocyclicin Q gene

August 19

Cloning

Digestion of PCR products (Including insert and backbone pET30a)
Ligation of digestion products (Ligase the DNA fragment into backbone pET30a)
Sample(8/17)

Enterocin B + pET30a
Enterocin 96 + pET30a
Bovicin HJ50 + pET30a
Durancin TW-49M + pET30a
Lacticin Z + pET30a
Leucocyclicin Q + pET30a

Sonication

Breaking E. coli ER2566 carrying the backbone pTXB1 and pTXB1 containing bacteriocin gene after cultivating to harvest the protein by sonication.
Sample

Culture of E. coli ER2566 carrying the backbone pTXB1 induced by IPTG
Culture of E. coli ER2566 carrying the backbone pTXB1 containing Enterocin B gene induced by IPTG
Culture of E. coli ER2566 carrying the backbone pTXB1 containing Enterocin 96 gene induced by IPTG
Culture of E. coli ER2566 carrying the backbone pTXB1 containing Leucocyclicin Q gene induced by IPTG

Protein quantification & SDS-PAGE

Quantificating the protein expressing the backbone pTXB1 and pTXB1 containing bacteriocin gene by Bradford method and running SDS-PAGE to check the protein expression.
Sample

Protein expressing the backbone pTXB1 induced by IPTG
Protein expressing the backbone pTXB1 containing Enterocin B gene induced by IPTG
Protein expressing the backbone pTXB1 containing Enterocin 96 gene induced by IPTG
Protein expressing the backbone pTXB1 containing Leucocyclicin Q gene induced by IPTG

August 20

Cloning

Transformation of ligation products (Transform into E. coli DH5α)
Taq PCR (PCR of ligation products to amplify insert gene)
Electrophoresis (To check PCR products)
Sample

Enterocin B + pET30a
Enterocin 96 + pET30a
Bovicin HJ50 + pET30a
Durancin TW-49M + pET30a
Lacticin Z + pET30a
Leucocyclicin Q + pET30a

August 21

Cloning

Cultivation
Miniprep (Purify Plasmid)
Sample

Enterocin B + pET30a
Enterocin 96 + pET30a
Bovicin HJ50 + pET30a
Durancin TW-49M + pET30a
Lacticin Z + pET30a
Leucocyclicin Q + pET30a

August 22

Cloning

Sequencing plasmid
Sample

Enterocin B + pET30a
Enterocin 96 + pET30a
Bovicin HJ50 + pET30a
Durancin TW-49M + pET30a
Lacticin Z + pET30a
Leucocyclicin Q + pET30a

August 23

None

August 24

Transformation

Transforming backbone pTXB1 containing bacteriocin gene into E. coli ER2566 to produce our target protein as a bio-stimulator.
Sample

backbone pTXB1 containing Bovicin HJ50 gene mini
backbone pTXB1 containing Durancin TW-49M gene mini

Cultivation

Cultivate the E. coli ER2566 at 37°C cultivating E. coli ER2566 at 37°C carrying the backbone pTXB1 containing bacteriocin gene to produce our target protein as a bio-stimulator.
Sample

Fresh colony of E. coli ER2566 carrying the backbone pTXB1 containing Bovicin HJ50 gene from an overnight plate
Fresh colony of E. coli ER2566 carrying the backbone pTXB1 containing Durancin TW-49M gene from an overnight plate

IPTG Induction

Inducting E. coli ER2566 carrying the backbone pTXB1 containing bacteriocin gene after cultivating at 37°C to produce our target protein as a bio-stimulator.
Sample

Culture of E. coli ER2566 carrying the backbone pTXB1 containing Bovicin HJ50 gene
Culture of E. coli ER2566 carrying the backbone pTXB1 containing Durancin TW-49M gene

August 25

Sonication

Breaking E. coli ER2566 carrying the backbone pTXB1 containing bacteriocin gene after cultivating to harvest the protein by sonication.
Sample

Culture of E. coli ER2566 carrying the backbone pTXB1 containing Bovicin HJ50 gene induced by IPTG
Culture of E. coli ER2566 carrying the backbone pTXB1 containing Durancin TW-49M gene induced by IPTG

Protein quantification & SDS-PAGE

Quantificating the protein expressing the backbone pTXB1 containing bacteriocin gene by Bradford method and running SDS-PAGE to check the protein expression.
Sample

Protein expressing the backbone pTXB1 containing Bovicin HJ50 gene induced by IPTG
Protein expressing the backbone pTXB1 containing Durancin TW-49M gene induced by IPTG

August 26

None

August 27

None

August 28

None

August 29

None

August 30

None

August 31

None

September 1

None

September 2

None

September 3

None

September 4

None

September 5

None

September 6

Transformation

Transforming backbone pTXB1 containing Lacticin Z gene into E. coli ER2566 to produce our target protein as a bio-stimulator.
Sample

backbone pTXB1 containing Lacticin Z gene mini

Cultivation

Cultivate the E. coli ER2566 at 37°C cultivating Bacillus subtilis ER2566 at 37°C carrying the backbone pTXB1 containing Lacticin Z gene to produce our target protein as a bio-stimulator.
Sample

Fresh colony of E. coli ER2566 carrying the backbone pTXB1 containing Lacticin Z gene from an overnight plate

IPTG Induction

Inducting E. coli ER2566 carrying the backbone pTXB1 containing Lacticin Z gene after cultivating at 37°C to produce our target protein as a bio-stimulator.
Sample

Culture of E. coli ER2566 carrying the backbone pTXB1 containing Lacticin Z gene

September 7

Sonication

Breaking E. coli ER2566 carrying the backbone pTXB1 containing Lacticin Z gene after cultivating to harvest the protein by sonication.
Sample

Culture of E. coli ER2566 carrying the backbone pTXB1 containing Lacticin Z gene induced by IPTG

Protein quantification & SDS-PAGE

Quantificating the protein expressing the backbone pTXB1 containing Lacticin Z gene by Bradford method and running SDS-PAGE to check the protein expression.
Sample

Protein expressing the backbone pTXB1 containing Lacticin Z gene induced by IPTG

September 8

None

September 9

None

September 10

None

September 11

None

September 12

None

September 13

None

September 14

None

September 15

None

September 16

None

September 17

None

September 18

None

September 19

None

September 20

None

September 21

Functional analysis test

Test the function of Lacticin Z (produced by E. coli ER2566) by elisa reader(triplicate)
Sample

Control:
1. Bacillus subtilis + Ampicillin
2. Bacillus subtilis + Production of the pTXB1 backbone induced by IPTG
3. Bacillus subtilis + LB broth
4. LB broth for background
Test:
1. Bacillus subtilis + Production of the pTXB1 backbone containing Lacticin Z gene induced by IPTG

September 22

None

September 23

None

September 24

None

September 25

None

September 26

None

September 27

None

September 28

None

September 29

None

September 30

None

October 1

None

October 2

None

October 3

None

October 4

None

October 5

Transformation

Transforming backbone pTXB1 into E. coli Rosetta-Gami to produce protein as our negative control in verifying the function of our target peptide.
Transforming backbone pTXB1 containing bacteriocin gene into E. coli Rosetta-Gami to produce our target protein as a bio-stimulator.
Sample

backbone pTXB1 mini
backbone pTXB1 containing Enterocin B gene mini
backbone pTXB1 containing Enterocin 96 gene mini
backbone pTXB1 containing Bovicin HJ50 gene mini
backbone pTXB1 containing Durancin TW-49M gene mini
backbone pTXB1 containing Lacticin Z gene mini
backbone pTXB1 containing Leucocyclicin Q gene mini

October 6

Cultivation

Cultivate the E. coli ER2566 at 37°C cultivating E. coli Rosetta-Gami at 37°C carrying the backbone pTXB1 to produce protein as our negative control in verifying the function of our target peptide.
Cultivate the E. coli ER2566 at 37°C cultivating E. coli Rosetta-Gami at 37°C carrying the backbone pTXB1 containing bacteriocin gene produce our target protein as a bio-stimulator.
Sample

Fresh colony of E. coli Rosetta-Gami carrying the backbone pTXB1 from an overnight plate
Fresh colony of E. coli Rosetta-Gami carrying the backbone pTXB1 containing Enterocin B gene from an overnight plate
Fresh colony of E. coli Rosetta-Gami carrying the backbone pTXB1 containing Enterocin 96 gene from an overnight plate
Fresh colony of E. coli Rosetta-Gami carrying the backbone pTXB1 containing Leucocyclicin Q gene from an overnight plate

IPTG Induction

Cultivate the E. coli ER2566 at 37°C cultivating E. coli Rosetta-Gami at 37°C carrying the backbone pTXB1 to produce protein as our negative control in verifying the function of our target peptide.
Inducting E. coli Rosetta-Gami carrying the backbone pTXB1 containing bacteriocin gene after cultivating at 37°C produce our target protein as a bio-stimulator.
Sample

Fresh colony of E. coli Rosetta-Gami carrying the backbone pTXB1 from an overnight plate
Culture of E. coli Rosetta-Gami carrying the backbone pTXB1 containing Enterocin B gene
Culture of E. coli Rosetta-Gami carrying the backbone pTXB1 containing Enterocin 96 gene
Culture of E. coli Rosetta-Gami carrying the backbone pTXB1 containing Leucocyclicin Q gene

October 7

Cultivation

Cultivate the E. coli ER2566 at 37°C cultivating E. coli Rosetta-Gami at 37°C carrying the backbone pTXB1 containing bacteriocin gene produce our target protein as a bio-stimulator.
Sample

Fresh colony of E. coli Rosetta-Gami carrying the backbone pTXB1 containing Bovicin HJ50 gene from an overnight plate
Fresh colony of E. coli Rosetta-Gami carrying the backbone pTXB1 containing Durancin TW-49M gene from an overnight plate
Fresh colony of E. coli Rosetta-Gami carrying the backbone pTXB1 containing Lacticin Z gene from an overnight plate

IPTG Induction

Inducting E. coli Rosetta-Gami carrying the backbone pTXB1 containing bacteriocin gene after cultivating at 37°C produce our target protein as a bio-stimulator.
Sample

Culture of E. coli Rosetta-Gami carrying the backbone pTXB1 containing Bovicin HJ50 gene
Culture of E. coli Rosetta-Gami carrying the backbone pTXB1 containing Durancin TW-49M gene
Culture of E. coli Rosetta-Gami carrying the backbone pTXB1 containing Lacticin Z gene

Sonication

Breaking E. coli Rosetta-Gami carrying the backbone pTXB1 and pTXB1 containing bacteriocin gene after cultivating to harvest the protein by sonication.
Sample

Culture of E. coli Rosetta-Gami carrying the backbone pTXB1 induced by IPTG
Culture of E. coli Rosetta-Gami carrying the backbone pTXB1 containing Enterocin B gene induced by IPTG
Culture of E. coli Rosetta-Gami carrying the backbone pTXB1 containing Enterocin 96 gene induced by IPTG
Culture of E. coli Rosetta-Gami carrying the backbone pTXB1 containing Leucocyclicin Q gene induced by IPTG

Protein quantification & SDS-PAGE

Quantificating the protein expressing the backbone pTXB1 and pTXB1 containing bacteriocin gene by Bradford method and running SDS-PAGE to check the protein expression.
Sample

Protein expressing the backbone pTXB1 induced by IPTG
Protein expressing the backbone pTXB1 containing Enterocin B gene induced by IPTG
Protein expressing the backbone pTXB1 containing Enterocin 96 gene induced by IPTG
Protein expressing the backbone pTXB1 containing Leucocyclicin Q gene induced by IPTG

October 8

Sonication

Breaking E. coli Rosetta-Gami carrying the backbone pTXB1 containing bacteriocin gene after cultivating to harvest the protein by sonication.
Sample

Culture of E. coli Rosetta-Gami carrying the backbone pTXB1 containing Bovicin HJ50 gene induced by IPTG
Culture of E. coli Rosetta-Gami carrying the backbone pTXB1 containing Durancin TW-49M gene induced by IPTG
Culture of E. coli Rosetta-Gami carrying the backbone pTXB1 containing Lacticin Z gene induced by IPTG
Culture of E. coli Rosetta-Gami carrying the backbone pTXB1 containing Leucocyclicin Q gene induced by IPTG

Protein quantification & SDS-PAGE

Quantificating the protein expressing the backbone pTXB1 containing bacteriocin gene by Bradford method and running SDS-PAGE to check the protein expression.
Sample

Protein expressing the backbone pTXB1 containing Bovicin HJ50 gene induced by IPTG
Protein expressing the backbone pTXB1 containing Durancin TW-49M gene induced by IPTG
Protein expressing the backbone pTXB1 containing Lacticin Z gene induced by IPTG
Protein expressing the backbone pTXB1 containing Leucocyclicin Q gene induced by IPTG

October 9

Functional analysis test

Test the function of bacteriocins (produced by E. coli Rosetta-gami) by elisa reader(triplicate)
Sample

Control:
1. Bacillus subtilis + Ampicillin
2. Bacillus subtilis + Production of the pTXB1 backbone induced by IPTG
3. Bacillus subtilis + LB broth
4. LB broth for background
Test:
1. Bacillus subtilis + Production of the pTXB1 backbone containing Enterocin B gene induced by IPTG
2. Bacillus subtilis + Production of the pTXB1 backbone containing Enterocin 96 gene induced by IPTG
3. Bacillus subtilis + Production of the pTXB1 backbone containing Lacticin Z gene induced by IPTG
4. Bacillus subtilis + Production of the pTXB1 backbone containing Durancin TW-49M gene induced by IPTG

October 10

Functional analysis test

Test the function of bacteriocins (produced by E. coli Rosetta-gami) by elisa reader(triplicate)
Sample

Control:
1. Bacillus subtilis + Ampicillin
2. Bacillus subtilis + Production of the pTXB1 backbone induced by IPTG
3. Bacillus subtilis + LB broth
4. LB broth for background
Test:
1. Bacillus subtilis + Production of the pTXB1 backbone containing Bovicin HJ50 gene induced by IPTG
2. Bacillus subtilis + Production of the pTXB1 backbone containing Leucocyclicin Q gene induced by IPTG

October 11

None

October 12

Functional analysis test

Test the function of bacteriocins (produced by E. coli Rosetta-gami) by elisa reader(Dose response assessment)
Sample

Control:
1. Bacillus subtilis + Ampicillin
2. Bacillus subtilis + Production of the pTXB1 backbone induced by IPTG
3. Bacillus subtilis + LB broth
4. LB broth for background
Test:
1. Bacillus subtilis + Production of the pTXB1 backbone containing Enterocin B gene induced by IPTG
2. Bacillus subtilis + Production of the pTXB1 backbone containing Enterocin 96 gene induced by IPTG
3. Bacillus subtilis + Production of the pTXB1 backbone containing Lacticin Z gene induced by IPTG
4. Bacillus subtilis + Production of the pTXB1 backbone containing Durancin TW-49M gene induced by IPTG

October 13

None

October 14

None

October 15

Functional analysis test

Test the function of bacteriocins (produced by E. coli Rosetta-gami) by elisa reader(Dose response assessment)
Sample

Control:
1. Bacillus subtilis + Ampicillin
2. Bacillus subtilis + Production of the pTXB1 backbone induced by IPTG
3. Bacillus subtilis + LB broth
4. LB broth for background
Test:
1. Bacillus subtilis + Production of the pTXB1 backbone containing Bovicin HJ50 gene induced by IPTG
2. Bacillus subtilis + Production of the pTXB1 backbone containing Leucocyclicin Q gene induced by IPTG

Functional analysis test(inhibition zone)

Test the function of Enterocin B (produced by E. coli Rosetta-gami) by LB Agar + Ampicillin plate
Sample

The Bacillus subtilis on the plate
Control:
1. Ampicillin
2. Production of the pTXB1 backbone induced by IPTG
Test:
1. Production of the pTXB1 backbone containing Enterocin B gene induced by IPTG

October 16

Functional analysis test(inhibition zone)

Test the function of bacteriocins (produced by E. coli Rosetta-gami) by LB Agar + Ampicillin plate
Sample

The Bacillus subtilis on the plate
Control:
1. Ampicillin
2. Production of the pTXB1 backbone induced by IPTG
Test:
1. Production of the pTXB1 backbone containing Bovicin HJ50 gene induced by IPTG
2. Production of the pTXB1 backbone containing Lacticin Z gene induced by IPTG

October 17

Functional analysis test

Test the function of bacteriocins (produced by E. coli Rosetta-gami) by elisa reader(Dose response assessment)
Sample

Control:
1. Bacillus subtilis + Ampicillin
2. Bacillus subtilis + Production of the pTXB1 backbone induced by IPTG
3. Bacillus subtilis + LB broth
Test:
1. Bacillus subtilis + Production of the pTXB1 backbone containing Durancin TW-49M gene induced by IPTG

October 18

None

October 19

None

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+    margin-left: 10%;
+    position: relative;
+    margin-top: 2vw;
+  }
 }
 .menu__wrapper {
 	padding: 6vmin;
    padding-top: 4vmin;
-   padding-left: 8vmin;
+   padding-left: 3vw;
-   padding-bottom: 0;
+   padding-bottom: 2vw;
 	position: relative;
 	z-index: 400;
@@ Line 311: / Line 321: @@
 th{
-    font-size:1.5em;
+    font-size:2.3vw;
     text-align:center;
     margin: 15px;
@@ Line 317: / Line 327: @@
 table tr td{
-    font-size:1.5em;
+    font-size:2.6vw;
     text-align:center;
@@ Line 334: / Line 344: @@
 .notebook{
-     margin-left:48vw;
+     margin-left:10%;
-     margin-top:-450px;
+     margin-bottom:150px;
-     margin-bottom:30px;
+    width: 35%;
+    display: inline-block;
+     margin-top: 3vw;
 }
 .notebook_wrapper{
      background:#b1e0f2;
-     width:510px;
+     width:100%;
      height:490px;
      border-radius:10px;
@@ Line 356: / Line 368: @@
      text-align:center;
      color: #b1e0f2;
-     font-size:3em;
+     font-size:4em;
-     padding: 10px;
+     padding: 13px;
      border-radius: 10px;
      background: #053B77;
@@ Line 380: / Line 392: @@
 .notebook_wrapper .note-item .note-title{
-     font-size:2em !important;
+     font-size:3em !important;
      font-weight:bold !important;
@@ Line 393: / Line 405: @@
      padding-bottom:8px;
      padding-left:5px !important;
+}
+.lab{
+  width: 15%;
+  position: absolute;
+  left: 73%;
+  top: 30vw;
+  transition-duration: 0.4s;
+}
+.protocol{
+  width: 15%;
+  position: absolute;
+  left: 73%;
+  top: 37vw;
+  transition-duration: 0.4s;
+}
+.lab:hover{
+  transform: scale(1.1);
+}
+.protocol:hover{
+  transform: scale(1.1);
 }
 </style>
 </head>
 <body>
      <div class="img-container">
          <img src="https://static.igem.org/mediawiki/2018/3/31/T--NCTU_Formosa--Notebook_cover.png" width="100%">
+        <a href="https://2018.igem.org/Team:NCTU_Formosa/Notebook"><img src="https://static.igem.org/mediawiki/2018/4/4c/T--NCTU_Formosa--Notebook_Lab_Note.png" class="lab"></a>
+        <a href="https://2018.igem.org/Team:NCTU_Formosa/Protocol"><img src="https://static.igem.org/mediawiki/2018/4/4f/T--NCTU_Formosa--Notebook_Lab_protocol.png" class="protocol"></a>
      </div>
 	<!-- Contenedor principal -->
@@ Line 669: / Line 708: @@
       <article id="July-1" class="note-item note_active">
          <h2>July 1</h2><hr>
-         <p class="note-title">None</p>
+         <p class="note-title">Cloning</p>
+        <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
+            <li class="list">Received and resuspended of the IDT DNA fragments</li>
+            <li class="list">Sample</li>
+                 <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
+                      <li class="list">GS linker+αS1-casein</li>
+                      <li class="list">Enterocin B</li>
+                      <li class="list">Enterocin 96</li>
+                      <li class="list">Bovicin HJ50</li>
+                      <li class="list">Durancin TW49M</li>
+                      <li class="list">Lacticin Z</li>
+                      <li class="list">Leucocyclicin Q</li>
+                 </ul>
+        </ul>
+        <p class="note-title"></p>
+        <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
+            <li class="list">Received and resuspended of the IDT DNA fragments</li>
+        </ul>
       </article>
       <article id="July-2" class="note-item">
@@ Line 677: / Line 733: @@
       <article id="July-3" class="note-item">
          <h2>July 3</h2><hr>
-         <p class="note-title">none</p>
+         <p class="note-title">Growth curve exp.</p>
+        <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
+            <li class="list">Observe the growth population of <i>Bacillus subtilis</i> in different temperature Condition: 37°C, 32°C, 27°C</li>
+        </ul>
       </article>
       <article id="July-4" class="note-item">
          <h2>July 4</h2><hr>
-         <p class="note-title">None</p>
+         <p class="note-title">Cloning gene of Curcumin bio-sensor</p>
+        <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
+            <li class="list">Doing the PCR and clean-up to amplify the DNA fragment. Digesting pET30a backbone and ligating it with αS1-casein+GS linker and T7 promoter.</li>
+            <li class="list">Sample</li>
+                 <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
+                      <li class="list">pET30a backbone</li>
+                      <li class="list">DNA segment of αS1-casein+GS linker and T7 promoter.</li>
+                 </ul>
+        </ul>
       </article>
       <article id="July-5" class="note-item">
          <h2>July 5</h2><hr>
-         <p class="note-title">Restore IDT in gblock fragment</p>
+         <p class="note-title">Cloning gene of Curcumin bio-sensor Expression</p>
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-             <li class="list">Save DNA</li>
+             <li class="list">Transforming the ligation product of the Curcumin bio-sensor into <i>E. coli</i> BL21 DE3 Cultivating the culture, preparing <i>E. coli</i> culture glycerol stocks, and doing the mini preparation.</li>
              <li class="list">Sample</li>
                   <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                       <li class="list">Hv1a-his-Sf1a NO.98351612</li>
+                       <li class="list">fresh colony of <i>E. coli</i> BL21 DE3 carrying the backbone pET30a containing αS1-casein + GS linker and T7 promotor gene from an overnight plate</li>
-                      <li class="list">Sf1a-his- OAIP NO.98351613</li>
-                      <li class="list">OAIP-his- Hv1a NO.98351614</li>
-                      <li class="list">Hv1a-lectin-his NO.98351615</li>
-                      <li class="list">Sf1a-lectin-his NO.98351616</li>
-                      <li class="list">OAIP-lectin-his NO.98351617</li>
                   </ul>
          </ul>
@@ Line 707: / Line 769: @@
       <article id="July-7" class="note-item">
          <h2>July 7</h2><hr>
-         <p class="note-title">None</p>
+         <p class="note-title">Expression of αS1-casein</p>
+        <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
+            <li class="list">Cultivating <i>E. coli</i> culture containing target protein αS1-casein from the <i>E. coli</i> glycerol stock and inducing it with IPTG to produce αS1-casein.</li>
+            <li class="list">Purifying the protein by His-Tag.</li>
+        </ul>
       </article>
       <article id="July-8" class="note-item">
          <h2>July 8</h2><hr>
-         <p class="note-title">None</p>
+         <p class="note-title">Growth curve exp</p>
+        <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
+            <li class="list">Observe the growth population of <i>Bacillus subtilis</i> in different pH Condition: pH = 4, 5, 6, 7, 8</li>
+        </ul>
       </article>
       <article id="July-9" class="note-item">
          <h2>July 9</h2><hr>
-         <p class="note-title">PCR of IDT gblock fragment</p>
+         <p class="note-title">None</p>
-           <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-               <li class="list">The IDT PCR amplify the insert gene and use phusion DNA polymerase (PFU) to improve the PCR accuracy rate. </li>
-               <li class="list">Sample</li>
-                 <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                      <li class="list">Hv1a-his-Sf1a</li>
-                      <li class="list">Sf1a-his- OAIP</li>
-                      <li class="list">OAIP-his- Hv1a</li>
-                      <li class="list">Hv1a-lectin-his</li>
-                      <li class="list">Sf1a-lectin-his</li>
-                      <li class="list">OAIP-lectin-his</li>
-                 </ul>
-        </ul>
-        <p class="note-title">Electrophoresis</p>
-           <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-               <li class="list">Check IDT PCR product</li>
-               <li class="list">Sample(7/9 PCR product)</li>
-                 <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                      <li class="list">Hv1a-his</li>
-                      <li class="list">Sf1a-his</li>
-                      <li class="list">OAIP-his</li>
-                      <li class="list">Hv1a-lectin-his</li>
-                      <li class="list">Sf1a-lectin-his</li>
-                      <li class="list">OAIP-lectin-his</li>
-                 </ul>
-        </ul>
       </article>
       <article id="July-10" class="note-item">
          <h2>July 10</h2><hr>
-         <p class="note-title">PCR of IDT gblock fragment</p>
+         <p class="note-title">Chip Production</p>
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-               <li class="list">Amplify the DNA to revise the mistake.</li>
+            <li class="list">Preparing 18 chips for bio-sensor sensitivity pretest<br>
-               <li class="list">Sample</li>
+. Dip the gold chips in 10mM Mua, RT for 4hrs.<br>
-                 <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
+. Wash the chips with 95% EtOH three times and dry.<br>
-                      <li class="list">Hv1a-his-Sf1a </li>
+. Add EDC+NHS mixture (100+100mM in DDW) on chips, RT for 1hrs.<br>
-                      <li class="list">Sf1a-his- OAIP</li>
+. DDW rinse the chips and dry.<br>
-                      <li class="list">OAIP-his- Hv1a </li>
+. Add αS1-casein on chips, RT for 1hrs.<br>
-                      <li class="list">Hv1a-lectin-his</li>
+. Wash with PBS three times and dry.<br>
-                      <li class="list">Sf1a-lectin-his</li>
+. Dip the chips in blocking solution, RT for 1.5hrs.<br>
-                      <li class="list">OAIP-lectin-his</li>
+. Wash with PBS three times and dry.</li>
-                 </ul>
-        </ul>
-        <p class="note-title">Electrophoresis</p>
-           <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-               <li class="list">Check IDT PCR product</li>
-               <li class="list">Sample(7/10 PCR product)</li>
-                 <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                      <li class="list">Hv1a-his</li>
-                      <li class="list">Sf1a-his</li>
-                      <li class="list">OAIP-his</li>
-                      <li class="list">Hv1a-lectin-his</li>
-                      <li class="list">Sf1a-lectin-his</li>
-                      <li class="list">OAIP-lectin-his</li>
-                 </ul>
-        </ul>
-        <p class="note-title">Gel extraction</p>
-           <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-               <li class="list">For the purification of DNA, we cut the correct electrophoresis band to do gel extraction.</li>
-               <li class="list">Sample</li>
-                 <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                      <li class="list">Hv1a-his</li>
-                      <li class="list">Sf1a-his</li>
-                      <li class="list">OAIP-his</li>
-                      <li class="list">Hv1a-lectin-his</li>
-                      <li class="list">Sf1a-lectin-his</li>
-                      <li class="list">OAIP-lectin-his</li>
-                 </ul>
-        </ul>
-       <p class="note-title">Digestion</p>
-           <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-               <li class="list">Digest the gene fragment(7/10) and pSB1C3 with EcoRI and PstI to ligase the gene fragment into the pSB1C3.</li>
-               <li class="list">Sample</li>
-                 <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                      <li class="list">Hv1a-lectin-his 4 7.10 (EP)</li>
-                      <li class="list">Sf1a-his 1 7.10 (EP)</li>
-                      <li class="list">OAIP-his 2 7.10 (EP)</li>
-                      <li class="list">OAIP-lectin-his 5 7.10 (EP)</li>
-                      <li class="list">Hv1a-his 3 7.10 (EP) </li>
-                      <li class="list">Sf1a-lectin-his 6 7.10 (EP)</li>
-                      <li class="list">pSB1C3 (EP)</li>
-                 </ul>
-        </ul>
-        <p class="note-title">Electrophoresis</p>
-           <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-               <li class="list">Electrophoresis of digested product to check the pSB1C3 digestion.</li>
-               <li class="list">Sample</li>
-                 <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                      <li class="list">pSB1C3 (EP)</li>
-                 </ul>
-        </ul>
-        <p class="note-title">Ligation</p>
-           <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-               <li class="list">Ligase DNA fragment into the pSB1C3</li>
-               <li class="list">Sample digestion product</li>
-               <li class="list">Sample</li>
-                 <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                      <li class="list">Hv1a-lectin-his 4 7.10 (EP)</li>
-                      <li class="list">Sf1a-his 1 7.10 (EP)</li>
-                      <li class="list">OAIP-his 2 7.10 (EP)</li>
-                      <li class="list">OAIP-lectin-his 5 7.10 (EP)</li>
-                      <li class="list">Hv1a-his 3 7.10 (EP)</li>
-                      <li class="list">Sf1a-lectin-his 6 7.10 (EP)</li>
-                      <li class="list">pSB1C3 (EP)</li>
-                 </ul>
          </ul>
       </article>
@@ Line 828: / Line 806: @@
       <article id="July-11" class="note-item">
          <h2>July 11</h2><hr>
-         <p class="note-title">Transformation:</p>
+         <p class="note-title">Bio-sensor sensitivity pretest </p>
-           <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
+        <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-               <li class="list">Transformation of ligated product to DH5α:</li>
+            <li class="list">Using DPV(Differential Pulse Voltammetry) to check whether our bio-sensor can detect curcumin.<br>
-               <li class="list">Sample</li>
+. Add the diluted curcumin samples on our bio-sensor to react for 30min.<br>
-                 <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
+. Rinse with wash buffer and dry the chips.<br>
-                      <li class="list">Sf1a-his 1 7.10 C<sup>r</sup></li>
+. Wash the reference and counter electrodes with DDW, and dry them.<br>
-                      <li class="list">Hv1a-lectin-his 4 7.10 C<sup>r</sup></li>
+. Set up the three electrodes system within electrocheFunctional analysisal cell.<br>
-                      <li class="list">OAIP-his 2 7.10 C<sup>r</sup></li>
+. Use the prototype of electrocheFunctional analysisal machine to measure the DPV method</li>
-                      <li class="list">OAIP-lectin-his 5 7.10 C<sup>r</sup></li>
-                      <li class="list">Hv1a-his 3 7.10 C<sup>r</sup></li>
-                      <li class="list">Sf1a-lectin-his 6 7.10 C<sup>r</sup></li>
-                 </ul>
          </ul>
       </article>
@@ Line 845: / Line 819: @@
       <article id="July-12" class="note-item">
          <h2>July 12</h2><hr>
-         <p class="note-title">Electrophoresis</p>
+         <p class="note-title">Bio-sensor sensitivity assay –Determine Standard Curve and Create the Formula</p>
-           <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
+         <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-               <li class="list">Electrophoresis of gel extraction product to check the concentration of gene to adjust the ligation protocol.</li>
+            <li class="list">Detecting the diluted standard curcumin samples by curcumin bio-sensor and made the standard curve and doing the polynomial curve fitting.</li>
-               <li class="list">Sample</li>
-                 <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                      <li class="list">Hv1a-his</li>
-                      <li class="list">Sf1a-his</li>
-                      <li class="list">OAIP-his</li>
-                      <li class="list">Hv1a-lectin-his</li>
-                      <li class="list">Sf1a-lectin-his</li>
-                      <li class="list">OAIP-lectin-his</li>
-                 </ul>
-         </ul>
-        <p class="note-title">Transformation:</p>
-           <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-               <li class="list">Transformation of ligated product 20µL to BL21 competent cell 50µL</li>
-               <li class="list">Sample</li>
-                 <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                      <li class="list">Sf1a-his 1 7.10 C<sup>r</sup></li>
-                      <li class="list">Hv1a-lectin-his 4 7.10 C<sup>r</sup></li>
-                      <li class="list">OAIP-his 2 7.10 C<sup>r</sup></li>
-                      <li class="list">OAIP-lectin-his 5 7.10 C<sup>r</sup></li>
-                      <li class="list">Hv1a-his 3 7.10 C<sup>r</sup></li>
-                      <li class="list">Sf1a-lectin-his 6 7.10 C<sup>r</sup></li>
-                 </ul>
          </ul>
       </article>
@@ Line 875: / Line 827: @@
       <article id="July-13" class="note-item">
          <h2>July 13</h2><hr>
-         <p class="note-title">PCR</p>
+         <p class="note-title">Bio-sensor sensitivity assay -Detecting the real Samples from Turmeric Root</p>
-           <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
+        <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-               <li class="list">PCR of ligated products to amplify the insert gene.</li>
+            <li class="list">Milling the turmeric root and dividing the powder into two groups. One of them was added with extraction buffer but not underwent the extraction protocol, and the other was added with extraction buffer but underwent the extraction process</li>
-               <li class="list">Sample</li>
-                 <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                      <li class="list">Sf1a-his C<sup>r</sup> 1~8</li>
-                      <li class="list">Hv1a-lectin-his C<sup>r</sup> 1~8</li>
-                      <li class="list">OAIP-his C<sup>r</sup> 1~8</li>
-                      <li class="list">OAIP-lectin-his C<sup>r</sup> 1~8</li>
-                      <li class="list">Hv1a-his C<sup>r</sup> 1~8</li>
-                      <li class="list">Sf1a-lectin-his C<sup>r</sup> 1~8</li>
-                 </ul>
          </ul>
       </article>
@@ Line 892: / Line 835: @@
       <article id="July-14" class="note-item">
          <h2>July 14</h2><hr>
-         <p class="note-title">PCR</p>
+         <p class="note-title">None</p>
-           <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-               <li class="list">PCR of ligated products to amplify the insert gene.</li>
-               <li class="list">Sample</li>
-                 <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                      <li class="list">Sf1a-his C<sup>r</sup> 1~4</li>
-                      <li class="list">Hv1a-lectin-his C<sup>r</sup> 1~4</li>
-                      <li class="list">OAIP-his C<sup>r</sup> 1~4</li>
-                      <li class="list">OAIP-lectin-his C<sup>r</sup> 1~4</li>
-                      <li class="list">Hv1a-his C<sup>r</sup> 1~4</li>
-                      <li class="list">Sf1a-lectin-his C<sup>r</sup> 1~4</li>
-                 </ul>
-        </ul>
-        <p class="note-title">Electrophoresis</p>
-           <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-               <li class="list">Electrophoresis of PCR products to check the insert gene base pair</li>
-               <li class="list">Sample</li>
-                 <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                      <li class="list">Hv1a-his C<sup>r</sup> 1~4</li>
-                      <li class="list">Sf1a-his C<sup>r</sup> 1~4</li>
-                      <li class="list">OAIP-his C<sup>r</sup> 1~4</li>
-                      <li class="list">Hv1a-lectin-his C<sup>r</sup> 1~4</li>
-                      <li class="list">Sf1a-lectin-his C<sup>r</sup> 1~4</li>
-                      <li class="list">OAIP-lectin-his C<sup>r</sup> 1~4</li>
-                 </ul>
-        </ul>
-        <p class="note-title">Cultivation</p>
-           <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-               <li class="list">Cultivation of ligated product.</li>
-               <li class="list">Sample</li>
-                 <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                      <li class="list">Sf1a-his C<sup>r</sup> 3</li>
-                      <li class="list">Hv1a-lectin-his C<sup>r</sup> 4</li>
-                      <li class="list">OAIP-his C<sup>r</sup> 1</li>
-                      <li class="list">OAIP-lectin-his C<sup>r</sup> 3</li>
-                      <li class="list">Hv1a-his C<sup>r</sup> 3</li>
-                      <li class="list">Sf1a-lectin-his C<sup>r</sup> 4</li>
-                 </ul>
-        </ul>
       </article>
       <article id="July-15" class="note-item">
          <h2>July 15</h2><hr>
-         <p class="note-title">PCR</p>
+         <p class="note-title">Expression</p>
             <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-               <li class="list">PCR of ligated products to amplify the insert gene.</li>
+            <li class="list">Preparing competent cell <i>E. coli</i> ER2566</li>
-               <li class="list">Sample</li>
+           </ul>
-                 <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                      <li class="list">Sf1a-his C<sup>r</sup> 1~8</li>
-                      <li class="list">Hv1a-lectin-his C<sup>r</sup> 1~8</li>
-                      <li class="list">Hv1a-his C<sup>r</sup> 1~6</li>
-                      <li class="list">Sf1a-lectin-his C<sup>r</sup> 1~3</li>
-                 </ul>
-        </ul>
-        <p class="note-title">Electrophoresis</p>
+        <p class="note-title">Growth curve exp</p>
             <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                <li class="list">Electrophoresis of PCR products to check the insert gene base pair</li>
+                <li class="list">Observe the growth population of <i>Bacillus subtilis</i> in different salinity condition: 0.17M, 0.25M, 0.5M, 0.75M, 1.0M</li>
-               <li class="list">Sample</li>
-                 <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                      <li class="list">Sf1a-his C<sup>r</sup> 1~8</li>
-                      <li class="list">Hv1a-lectin-his C<sup>r</sup> 1~8</li>
-                      <li class="list">Hv1a-his C<sup>r</sup> 1~6</li>
-                      <li class="list">Sf1a-lectin-his C<sup>r</sup> 1~3</li>
-                 </ul>
-        </ul>
-       <p class="note-title">Transformation</p>
-           <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-               <li class="list">Transformation of Hv1a-lectin-his C<sup>r</sup>, Hv1a-his C<sup>r</sup> into BL21 again.</li>
             </ul>
-       <p class="note-title">Miniprep</p>
-           <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-               <li class="list">Purify plasmid which have insert gene.(7/14)</li>
-               <li class="list">Sample</li>
-                 <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                      <li class="list">Hv1a-his C<sup>r</sup></li>
-                      <li class="list">Sf1a-his C<sup>r</sup></li>
-                      <li class="list">OAIP-his C<sup>r</sup></li>
-                     <li class="list">Hv1a-lectin-his C<sup>r</sup></li>
-                      <li class="list">Sf1a-lectin-his C<sup>r</sup></li>
-                     <li class="list">OAIP-lectin-his C<sup>r</sup></li>
-                 </ul>
-        </ul>
       </article>
       <article id="July-16" class="note-item">
          <h2>July 16</h2><hr>
-         <p class="note-title">Digestion</p>
+         <p class="note-title">Cloning</p>
             <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                <li class="list">Digest the plasmid (7/15) with EcoRI and PstI to check whether the mini have insert gene fragment.</li>
+                <li class="list">Amplify the insert gene (Pfu PCR)</li>
+               <li class="list">Electrophoresis (To check PCR products)</li>
                 <li class="list">Sample</li>
                   <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                       <li class="list">Sf1a-his 7.15 (EP)</li>
+                       <li class="list">Lacticin Z + pTXB1</li>
-                       <li class="list">OAIP-his 7.15 (EP)</li>
+                       <li class="list">Bovincin HJ50+ pTXB1</li>
-                      <li class="list">Hv1a-his 7.15 (EP)</li>
-                      <li class="list">Sf1a-lectin-his 7.15 (EP)</li>
-                      <li class="list">OAIP-lectin-his 7.15 (EP)</li>
-                      <li class="list">Hv1a-lectin-his 7.15 (EP)</li>
                   </ul>
-        </ul>
+            </ul>
-               <p class="note-title">Electrophoresis</p>
-            <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-               <li class="list">Electrophoresis of digested product to check the band whether proper to the really base pair.</li>
-               <li class="list">Sample(7/16 Digestion product)</li>
-                 <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                      <li class="list">Sf1a-his 7.15 (EP)</li>
-                      <li class="list">OAIP-his 7.15 (EP)</li>
-                      <li class="list">Hv1a-his 7.15 (EP)</li>
-                      <li class="list">Sf1a-lectin-his 7.15 (EP)</li>
-                      <li class="list">OAIP-lectin-his 7.15 (EP)</li>
-                      <li class="list">Hv1a-lectin-his 7.15 (EP)</li>
-                 </ul>
-        </ul>
-       <p class="note-title">PCR</p>
-           <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-               <li class="list">PCR of ligated products to amplify the insert gene.</li>
-               <li class="list">Sample</li>
-                 <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                      <li class="list">Hv1a-his C<sup>r</sup> 1~11</li>
-                      <li class="list">Hv1a-lectin-his C<sup>r</sup> 1~7</li>
-                 </ul>
-        </ul>
-       <p class="note-title">Electrophoresis</p>
-           <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-               <li class="list">Electrophoresis of PCR products to check the insert gene base pair.</li>
-               <li class="list">Sample</li>
-                 <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                      <li class="list">Hv1a-his C<sup>r</sup> 1~11</li>
-                      <li class="list">Hv1a-lectin-his C<sup>r</sup> 1~7</li>
-                 </ul>
-        </ul>
-       <p class="note-title">Cultivation</p>
-           <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-               <li class="list">Cultivation of ligated product.</li>
-               <li class="list">Sample</li>
-                 <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                      <li class="list">Sf1a-his C<sup>r</sup> 1</li>
-                      <li class="list">Hv1a-his C<sup>r</sup> 6</li>
-                      <li class="list">Hv1a-lectin-his C<sup>r</sup> 1</li>
-                      <li class="list">Hv1a-lectin-his C<sup>r</sup> 8</li>
-                      <li class="list">Sf1a-lectin-his C<sup>r</sup> 1</li>
-                 </ul>
-        </ul>
       </article>
       <article id="July-17" class="note-item">
          <h2>July 17</h2><hr>
-         <p class="note-title">Digestion</p>
+         <p class="note-title">Cloning</p>
             <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                <li class="list">Digest the plasmid (7/15) with EcoRI and PstI to check whether the mini have insert gene fragment.</li>
+                <li class="list">Digestion of PCR products (Including insert and backbone pTXB1)</li>
+               <li class="list">Ligation of digestion products (Ligase the DNA fragment into backbone pTXB1)</li>
                 <li class="list">Sample</li>
                   <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                       <li class="list">Sf1a-his 7.15 (EP)</li>
+                       <li class="list">Lacticin Z + pTXB1</li>
-                       <li class="list">Hv1a-his 7.15 (EP)</li>
+                       <li class="list">Bovincin HJ50+ pTXB1</li>
-                      <li class="list">Sf1a-lectin-his 7.15 (EP)</li>
-                      <li class="list">Hv1a-lectin-his 7.15 (EP)</li>
                   </ul>
-        </ul>
+            </ul>
-               <p class="note-title">Electrophoresis</p>
-            <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-               <li class="list">Electrophoresis of digested product to check the band whether proper to the really base pair.</li>
-               <li class="list">Sample</li>
-                 <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                      <li class="list">Sf1a-his 7.15 (EP)</li>
-                      <li class="list">Hv1a-his 7.15 (EP)</li>
-                      <li class="list">Sf1a-lectin-his 7.15 (EP)</li>
-                      <li class="list">Hv1a-lectin-his 7.15 (EP)</li>
-                 </ul>
-        </ul>
-       <p class="note-title">Miniprep</p>
-           <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-               <li class="list">Purify plasmid which have insert gene.(7/16)</li>
-               <li class="list">Sample</li>
-                 <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                      <li class="list">Sf1a-his C<sup>r</sup> 1</li>
-                      <li class="list">Sf1a-lectin-his C<sup>r</sup> 1</li>
-                      <li class="list">Hv1a-his C<sup>r</sup> 6</li>
-                      <li class="list">Hv1a-lectin-his C<sup>r</sup> 1</li>
-                      <li class="list">Hv1a-lectin-his C<sup>r</sup> 8</li>
-                 </ul>
-        </ul>
-       <p class="note-title">Cultivation</p>
-           <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-               <li class="list">Cultivation of ligated product.(7/16)</li>
-               <li class="list">Sample</li>
-                 <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                      <li class="list">Hv1a-his C<sup>r</sup> 5</li>
-                      <li class="list">Hv1a-his C<sup>r</sup> 7</li>
-                      <li class="list">Hv1a-his C<sup>r</sup> 10</li>
-                      <li class="list">Hv1a-his C<sup>r</sup> 11</li>
-                      <li class="list">Hv1a-lectin-his C<sup>r</sup> 2</li>
-                      <li class="list">Hv1a-lectin-his C<sup>r</sup> 3</li>
-                      <li class="list">Hv1a-lectin-his C<sup>r</sup> 7</li>
-                 </ul>
-        </ul>
       </article>
       <article id="July-18" class="note-item">
          <h2>July 18</h2><hr>
-        <p class="note-title">PRC of IDT gblock fragment:</p>
+        <p class="note-title">Cloning</p>
             <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                <li class="list">Amplify the DNA to revise the mistake.</li>
+                <li class="list">Transformation of ligation products (Transform into <i>E. coli</i> DH5α)</li>
+               <li class="list">Taq PCR (PCR of ligation products to amplify insert gene)</li>
+               <li class="list">Electrophoresis (To check PCR products)</li>
                 <li class="list">Sample</li>
                   <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                       <li class="list">Hv1a-his-Sf1a NO.98351612</li>
+                       <li class="list">Lacticin Z + pTXB1</li>
-                       <li class="list">Sf1a-his-OAIP NO.98351613</li>
+                       <li class="list">Bovincin HJ50+ pTXB1</li>
-                      <li class="list">OAIP-his-Hv1a NO.98351614</li>
-                      <li class="list">Hv1a-lectin-his NO.98351615</li>
-                      <li class="list">Sf1a-lectin-his NO.98351616</li>
-                      <li class="list">OAIP-lectin-his NO.98351617</li>
                   </ul>
-        </ul>
+            </ul>
-        <p class="note-title">Miniprep</p>
-            <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-               <li class="list">Purify plasmid which have insert gene.(7/17)</li>
-               <li class="list">Sample</li>
-                 <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                      <li class="list">Hv1a-his C<sup>r</sup> 5</li>
-                      <li class="list">Hv1a-his C<sup>r</sup> 7</li>
-                      <li class="list">Hv1a-his C<sup>r</sup> 10</li>
-                      <li class="list">Hv1a-his C<sup>r</sup> 11</li>
-                      <li class="list">Hv1a-lectin-his C<sup>r</sup> 3</li>
-                      <li class="list">Hv1a-lectin-his C<sup>r</sup> 7</li>
-                 </ul>
-        </ul>
-        <p class="note-title">Cultivation</p>
-           <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-               <li class="list">Cultivation of ligated product.(7/17)</li>
-               <li class="list">Sample</li>
-                 <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                      <li class="list">OAIP-lectin-his C<sup>r</sup> 1</li>
-                      <li class="list">OAIP-lectin-his C<sup>r</sup> 2</li>
-                 </ul>
-        </ul>
-       <p class="note-title">Digestion</p>
-           <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-               <li class="list">Digest the plasmid (7/18) with EcoRI and PstI to check whether the mini have insert gene fragment.</li>
-               <li class="list">Sample</li>
-                 <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                      <li class="list">Hv1a-his(EP)</li>
-                      <li class="list">Hv1a-his(EP)</li>
-                      <li class="list">Hv1a-his(EP)</li>
-                      <li class="list">Hv1a-his(EP)</li>
-                      <li class="list">Hv1a-lectin-his(EP)</li>
-                      <li class="list">Hv1a-lectin-his(EP)</li>
-                 </ul>
-        </ul>
-       <p class="note-title">Gel Electrophoresis of PCR-Amplified gBlocks</p>
-           <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-               <li class="list"></li>
-               <li class="list">Sample</li>
-                 <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                      <li class="list">Hv1a-his</li>
-                      <li class="list">Sf1a-his</li>
-                      <li class="list">OAIP-his</li>
-                      <li class="list">Hv1a-lectin-his</li>
-                      <li class="list">sf1a-lectin-his</li>
-                      <li class="list">OAIP-lectin-his</li>
-                 </ul>
-        </ul>
-       <p class="note-title">Digestion</p>
-           <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-               <li class="list">Digest the gene fragment(7/18) and pSB1A3 with EcoRI and PstI to ligase the gene fragment into the pSB1A3.</li>
-               <li class="list">Sample</li>
-                 <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                      <li class="list">Sf1a-his 1 7.18 (EP)</li>
-                      <li class="list">Hv1a-lectin-his 4 7.18 (EP)</li>
-                      <li class="list">OAIP-his 2 7.18 (EP)</li>
-                      <li class="list">OAIP-lectin-his 5 7.18 (EP)</li>
-                      <li class="list">Hv1a-his 3 7.18 (EP)</li>
-                      <li class="list">Sf1a-lectin-his 6 7.18 (EP)</li>
-                      <li class="list">pSB1C3 (EP)</li>
-                 </ul>
-        </ul>
-       <p class="note-title">Electrophoresis</p>
-           <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-               <li class="list">Electrophoresis of digested product to check the pSB1C3 digestion.</li>
-               <li class="list">Sample7/18 Digestion product</li>
-        </ul>
-       <p class="note-title">Ligation</p>
-           <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-               <li class="list">Ligase DNA fragment(7/18) into the pSB1C3</li>
-               <li class="list">Sample</li>
-                 <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                      <li class="list">Sf1a-his 1 7.18 (EP)</li>
-                      <li class="list">Hv1a-lectin-his 4 7.18 (EP)</li>
-                      <li class="list">OAIP-his 2 7.18 (EP)</li>
-                      <li class="list">OAIP-lectin-his 5 7.18 (EP)</li>
-                      <li class="list">Hv1a-his 3 7.18 (EP)</li>
-                      <li class="list">Sf1a-lectin-his 6 7.18 (EP)</li>
-                      <li class="list">pSB1C3 (EP)</li>
-                 </ul>
-        </ul>
-       <p class="note-title">Electrophoresis</p>
-           <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-               <li class="list">check the ligation product.</li>
-               <li class="list">Sample7/18 Ligation product</li>
-        </ul>
       </article>
       <article id="July-19" class="note-item">
          <h2>July 19</h2><hr>
-        <p class="note-title">Electrophoresis</p>
+        <p class="note-title">Cloning</p>
             <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                <li class="list">Electrophoresis of ligated product.</li>
+                <li class="list">Cultivation</li>
+               <li class="list">Miniprep (Purify Plasmid)</li>
                 <li class="list">Sample</li>
                   <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                       <li class="list">Hv1a-his</li>
+                       <li class="list">Lacticin Z + pTXB1</li>
-                       <li class="list">Sf1a-his</li>
+                       <li class="list">Bovincin HJ50+ pTXB1</li>
-                      <li class="list">OAIP-his</li>
-                      <li class="list">Hv1a-lectin-his</li>
-                      <li class="list">Sf1a-lectin-his</li>
-                      <li class="list">OAIP-lectin-his</li>
                   </ul>
-        </ul>
+            </ul>
-        <p class="note-title">Transformation</p>
-            <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-               <li class="list">Transformation of ligated product to DH5α.</li>
-               <li class="list">Sample</li>
-                 <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                      <li class="list">Hv1a-his 7/18 A<sup>r</sup></li>
-                      <li class="list">Sf1a-his 7/18 A<sup>r</sup></li>
-                      <li class="list">OAIP-his 7/8 A<sup>r</sup></li>
-                      <li class="list">Hv1a-lectin-his 7/8 A<sup>r</sup></li>
-                      <li class="list">sf1a-lectin-his 7/8 A<sup>r</sup></li>
-                      <li class="list">OAIP-lectin-his 7/18 A<sup>r</sup></li>
-                 </ul>
-        </ul>
-        <p class="note-title">Miniprep</p>
-           <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-               <li class="list">Purify plasmid which have insert gene.(7/18)</li>
-               <li class="list">Sample</li>
-                 <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                      <li class="list">OAIP-lectin-his C<sup>r</sup> 7/19 1</li>
-                      <li class="list">OAIP-lectin-his C<sup>r</sup> 7/19 2</li>
-                 </ul>
-        </ul>
-       <p class="note-title">Digestion</p>
-           <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-               <li class="list">Digest the plasmid (7/19) with EcoRI and PstI to check whether the mini have insert gene fragment.</li>
-               <li class="list">Sample</li>
-                 <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                      <li class="list">Hv1a-lectin-his C<sup>r</sup> 1 (EP)</li>
-                      <li class="list">Hv1a-lectin-his C<sup>r</sup> 2 (EP)</li>
-                 </ul>
-        </ul>
-       <p class="note-title">Electrophoresis</p>
-           <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-               <li class="list">Electrophoresis of ligated product.</li>
-               <li class="list">Sample</li>
-                 <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                      <li class="list">Hv1a-lectin-his C<sup>r</sup> 7/19 1</li>
-                      <li class="list">Hv1a-lectin-his C<sup>r</sup> 7/19 2</li>
-                 </ul>
-        </ul>
-       <p class="note-title">Digestion</p>
-           <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-               <li class="list">Digest the plasmid (7/18) with EcoRI and PstI to check whether the mini have insert gene fragment.</li>
-               <li class="list">Sample</li>
-                 <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                      <li class="list">Hv1a-his C<sup>r</sup> 1</li>
-                      <li class="list">Hv1a-his C<sup>r</sup> 2</li>
-                      <li class="list">Hv1a-his C<sup>r</sup> 3</li>
-                      <li class="list">Hv1a-his C<sup>r</sup> 4</li>
-                      <li class="list">Hv1a-lectin-his C<sup>r</sup> 1</li>
-                      <li class="list">Hv1a-lectin-his C<sup>r</sup> 2</li>
-                 </ul>
-        </ul>
-       <p class="note-title">Electrophoresis</p>
-           <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-               <li class="list">Electrophoresis of digestion product.</li>
-               <li class="list">Sample</li>
-                 <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                      <li class="list">Hv1a-his C<sup>r</sup> 1</li>
-                      <li class="list">Hv1a-his C<sup>r</sup> 2</li>
-                      <li class="list">Hv1a-his C<sup>r</sup> 3</li>
-                      <li class="list">Hv1a-his C<sup>r</sup> 4</li>
-                      <li class="list">Hv1a-lectin-his C<sup>r</sup> 1</li>
-                      <li class="list">Hv1a-lectin-his C<sup>r</sup> 2</li>
-                 </ul>
-        </ul>
       </article>
       <article id="July-20" class="note-item">
          <h2>July 20</h2><hr>
-        <p class="note-title">PCR</p>
+        <p class="note-title">Cloning</p>
             <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                <li class="list">PCR of ligated products(7/19) to amplify the insert gene.</li>
+                <li class="list">Sequencing plasmid</li>
                 <li class="list">Sample</li>
                   <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                       <li class="list">Hv1a-his A<sup>r</sup> 1~4</li>
+                       <li class="list">Lacticin Z + pTXB1</li>
-                       <li class="list">Sf1a-his A<sup>r</sup> 1~4</li>
+                       <li class="list">Bovincin HJ50+ pTXB1</li>
-                      <li class="list">OAIP-his A<sup>r</sup> 1~4</li>
-                      <li class="list">Hv1a-lectin-his A<sup>r</sup> 1~4</li>
-                      <li class="list">Sf1a-lectin-his A<sup>r</sup> 1~4</li>
-                      <li class="list">OAIP-lectin-his A<sup>r</sup> 1~4</li>
-                 </ul>
-        </ul>
-        <p class="note-title">Digestion</p>
-           <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-               <li class="list">Digest the gene fragment(7/18) and pSB1A3 with EcoRI and PstI to ligase the gene fragment into the pSB1A3.</li>
-               <li class="list">Sample</li>
-                 <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                      <li class="list">Hv1a-his 7/18 C<sup>r</sup>(EP)</li>
-                      <li class="list">Hv1a-his 7/18 C<sup>r</sup>(EP)</li>
-                      <li class="list">Hv1a-his 7/18 C<sup>r</sup>(EP)</li>
-                      <li class="list">Hv1a-his 7/18 C<sup>r</sup>(EP)</li>
-                      <li class="list">Hv1a-lectin-his 7/18 C<sup>r</sup>(EP)</li>
-                      <li class="list">Hv1a-lectin-his 7/18 C<sup>r</sup>(EP)</li>
-                      <li class="list">OAIP-lectin-his 7/18 C<sup>r</sup>(EP)</li>
-                 </ul>
-        </ul>
-        <p class="note-title">Electrophoresis</p>
-           <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-               <li class="list">Electrophoresis of digestion product.</li>
-               <li class="list">Sample</li>
-                 <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                      <li class="list">Hv1a-his 1~4 7/18(EP)</li>
-                      <li class="list">Hv1a-lectin-his 1, 2 7/18 (EP)</li>
-                      <li class="list">OAIP-lectin-his 7/19(EP)</li>
-                      <li class="list">Hv1a-his 3 7/18</li>
-                      <li class="list">Hv1a-lectin-his 2 7/18</li>
-                      <li class="list"></li>
-                 </ul>
-        </ul>
-       <p class="note-title">Electrophoresis</p>
-           <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-               <li class="list">Check insert gene in E.coli.</li>
-               <li class="list">Sample</li>
-                 <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                      <li class="list">Hv1a-his 1-1~4</li>
-                      <li class="list">Hv1a-lectin-his 2-1~4</li>
-                      <li class="list">Sf1a-his3-1~4</li>
-                      <li class="list">Sf1a-lectin-his 4-1~4</li>
-                      <li class="list">OAIP-his 5-1~4</li>
-                      <li class="list">OAIP-lectin-his 6-1~4</li>
-                 </ul>
-        </ul>
-       <p class="note-title">Sequencing</p>
-           <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-               <li class="list">determine insert gene’s Sequence. </li>
-               <li class="list">Sample</li>
-                 <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                      <li class="list">Hv1a-his 3(The result is correct)</li>
-                      <li class="list">Hv1a-lectin-his 2</li>
-                      <li class="list">OAIP-lectin-his 1</li>
-                 </ul>
-        </ul>
-       <p class="note-title">Ligation</p>
-           <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-               <li class="list">Ligase DNA fragment(7/18) into the pSB1A3</li>
-               <li class="list">Sample digestion product</li>
-                 <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                      <li class="list">Hv1a-his 7/20 (EP)</li>
-                      <li class="list">Hv1a-lectin-his 7/20 (EP)</li>
-                      <li class="list">OAIP-lectin-his 7/20 (EP)</li>
                   </ul>
          </ul>
@@ Line 1,385: / Line 922: @@
       <article id="July-21" class="note-item">
          <h2>July 21</h2><hr>
-        <p class="note-title">Cultivation</p>
+        <p class="note-title">None</p>
-           <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
+     </article>
-               <li class="list">Cultivation of ligated product.(7/20)</li>
-               <li class="list">Sample</li>
-                 <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                      <li class="list">Hv1a-his A<sup>r</sup> 2~4</li>
-                      <li class="list">Sf1a-his A<sup>r</sup> 1~4</li>
-                      <li class="list">OAIP-his A<sup>r</sup> 1.3.4</li>
-                      <li class="list">Hv1a-lectin-his A<sup>r</sup> 1~4</li>
-                      <li class="list">Sf1a-lectin-his A<sup>r</sup> 1~4</li>
-                      <li class="list">OAIP-lectin-his A<sup>r</sup> 1.3.4</li>
-                 </ul>
-        </ul>
-        <p class="note-title">Miniprep</p>
-           <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-               <li class="list">Purify plasmid which have insert gene.(7/21)</li>
-               <li class="list">Sample</li>
-                 <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                      <li class="list">Hv1a-his A<sup>r</sup> 2~4</li>
-                      <li class="list">Sf1a-his A<sup>r</sup> 1~4</li>
-                      <li class="list">OAIP-his A<sup>r</sup> 1.3.4</li>
-                      <li class="list">Hv1a-lectin-his A<sup>r</sup> 1~4</li>
-                      <li class="list">Sf1a-lectin-his A<sup>r</sup> 1~4</li>
-                      <li class="list">OAIP-lectin-his A<sup>r</sup> 1.3.4</li>
-                 </ul>
-        </ul>
-        <p class="note-title">Transformation</p>
-           <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-               <li class="list">Transformation of ligated product(7/20) to <i>BL21-rosetta-gami</i></li>
-               <li class="list">Sample</li>
-                 <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                      <li class="list">Hv1a-his A<sup>r</sup></li>
-                      <li class="list">Hv1a-lectin-his A<sup>r</sup></li>
-                      <li class="list">OAIP-lectin-his A<sup>r</sup></li>
-                 </ul>
-        </ul>
-     </article>
       <article id="July-22" class="note-item">
          <h2>July 22</h2><hr>
-        <p class="note-title">Digestion</p>
+        <p class="note-title">Cloning</p>
             <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                <li class="list">Digest the plasmid (7/21) with EcoRI and PstI to check whether the mini have insert gene fragment.</li>
+                <li class="list">Amplify the insert gene (Pfu PCR)</li>
+               <li class="list">Electrophoresis (To check PCR products)</li>
+               <li class="list">Digestion of PCR products (Including insert and backbone pTXB1)</li>
+               <li class="list">Ligation of digestion products (Ligase the DNA fragment into backbone pTXB1)</li>
                 <li class="list">Sample</li>
                   <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                       <li class="list">Hv1a-his A<sup>r</sup> 2~4</li>
+                       <li class="list">Leucocyclicin Q + pTXB1</li>
-                      <li class="list">Sf1a-his A<sup>r</sup> 1~4</li>
-                      <li class="list">OAIP-his A<sup>r</sup> 1.3.4</li>
-                      <li class="list">Hv1a-lectin-his A<sup>r</sup> 1~4</li>
-                      <li class="list">Sf1a-lectin-his A<sup>r</sup> 1~4</li>
-                      <li class="list">OAIP-lectin-his A<sup>r</sup> 1.3.4</li>
                   </ul>
-        </ul>
-        <p class="note-title">Electrophoresis</p>
-           <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-               <li class="list">Electrophoresis of ligated product.</li>
-               <li class="list">Sample</li>
-                 <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                      <li class="list">Hv1a-his A<sup>r</sup> 2~4</li>
-                      <li class="list">Sf1a-his A<sup>r</sup> 1~4</li>
-                      <li class="list">OAIP-his A<sup>r</sup> 1.3.4</li>
-                      <li class="list">Hv1a-lectin-his A<sup>r</sup> 1~4</li>
-                      <li class="list">Sf1a-lectin-his A<sup>r</sup> 1~4</li>
-                      <li class="list">OAIP-lectin-his A<sup>r</sup> 1.3.4</li>
-                 </ul>
-        </ul>
-        <p class="note-title">Sequencing</p>
-           <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-               <li class="list">determine insert gene’s Sequence.</li>
-               <li class="list">Sample</li>
-                 <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                      <li class="list">Hv1a-his (The result is correct)</li>
-                      <li class="list">Hv1a -his (The result is correct)</li>
-                      <li class="list">OAIP -his (The result is correct)</li>
-                 </ul>
-        </ul>
-       <p class="note-title">Digestion</p>
-           <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-               <li class="list">Digest the plasmid (7/21) with EcoRI and PstI to check whether the mini have insert gene fragment.</li>
-               <li class="list">Sample</li>
-                 <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                      <li class="list">Sf1a-his A<sup>r</sup> 1~4(EP)</li>
-                      <li class="list">OAIP-his A<sup>r</sup> 1.3.4(EP)</li>
-                      <li class="list">OAIP-lectin-his A<sup>r</sup> 1.3.4(EP)</li>
-                 </ul>
-        </ul>
-       <p class="note-title">Digestion</p>
-           <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-               <li class="list">Digest IDT gBlock fragment for ES because lectin have P Cut position.</li>
-               <li class="list">Sample</li>
-                 <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                      <li class="list">Hv1a-lectin-his</li>
-                      <li class="list">OAIP-lectin-his</li>
-                      <li class="list">Sf1a-lectin-his</li>
-                      <li class="list">Psb1A3</li>
-                 </ul>
-        </ul>
-       <p class="note-title">Electrophoresis</p>
-           <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-               <li class="list">Electrophoresis of ligated product.</li>
-               <li class="list">Sample digestion product</li>
-                 <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                      <li class="list">Sf1a-his A<sup>r</sup> 1~4(EP)</li>
-                      <li class="list">OAIP-his A<sup>r</sup> 1.3.4(EP)</li>
-                      <li class="list">OAIP-lectin-his A<sup>r</sup> 1.3.4(EP)</li>
-                 </ul>
-        </ul>
-       <p class="note-title">PRC of IDT gblock fragment</p>
-           <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-               <li class="list">Amplify the DNA to revise the mistake.</li>
-               <li class="list">Sample</li>
-                 <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                      <li class="list">Hv1a-his</li>
-                      <li class="list">Sf1a -his</li>
-                      <li class="list">OAIP-his</li>
-                      <li class="list">Hv1a-lectin-his</li>
-                      <li class="list">Sf1a-lectin-his</li>
-                      <li class="list">OAIP-lectin-his</li>
-                 </ul>
-        </ul>
-       <p class="note-title">Electrophoresis</p>
-           <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-               <li class="list">Electrophoresis of digested product to check insert gene.</li>
-               <li class="list">Sample</li>
-                 <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                      <li class="list">Hv1a-lectin-his (ES)</li>
-                      <li class="list">Sf1a-lectin-his (ES)</li>
-                      <li class="list">OAIP-lectin-his (ES)</li>
-                      <li class="list">[pSB1A3] (ES)</li>
-                 </ul>
-        </ul>
-       <p class="note-title">Ligation</p>
-           <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-               <li class="list">Ligase DNA fragment(7/18) into the pSB1A3</li>
-               <li class="list">Sample</li>
-                 <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                      <li class="list">Hv1a-lectin-his (ES)</li>
-                      <li class="list">Sf1a-lectin-his (ES)</li>
-                      <li class="list">OAIP-lectin-his (ES)</li>
-                      <li class="list">[pSB1A3] (ES)</li>
-                 </ul>
-        </ul>
-       <p class="note-title">Transformation</p>
-           <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-               <li class="list">Transformation of backbone plasmid into <i>BL21-rosetta-gami</i> check whether the competent cell is working.</li>
-               <li class="list">we choose pSB1A3 to transformation because <i>BL21-rosetta-gami</i> has C<sup>r</sup> and K<sup>r</sup>.</li>
          </ul>
       </article>
@@ Line 1,551: / Line 942: @@
       <article id="July-23" class="note-item">
          <h2>July 23</h2><hr>
-        <p class="note-title">Electrophoresis</p>
+        <p class="note-title">Cloning</p>
             <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                <li class="list">After the PRC of IDT gBlock fragment, we electrophoresis samples to check the insert gene fragment.</li>
+                <li class="list">Transformation of ligation products (Transform into <i>E. coli</i> DH5α)</li>
+               <li class="list">Taq PCR (PCR of ligation products to amplify insert gene)</li>
+               <li class="list">Electrophoresis (To check PCR products)</li>
                 <li class="list">Sample</li>
                   <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                       <li class="list">Hv1a-his</li>
+                       <li class="list">Leucocyclicin Q + pTXB1</li>
-                      <li class="list">Sf1a-his</li>
-                      <li class="list">OAIP-his</li>
-                      <li class="list">Hv1a-lectin-his</li>
-                      <li class="list">Sf1a-lectin-his</li>
-                      <li class="list">OAIP-lectin-his</li>
-                 </ul>
-        </ul>
-        <p class="note-title">Digestion</p>
-           <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-               <li class="list">There is a fragment in the snow drop lectin identification with PstI. Therefore, if sequence have snow drop lectin, we digested the gene fragment(7/22) and pSB1A3 with EcoRI and Spe l to ligase the gene fragment into the pSB1A3. Others still digested with EcoRI and PstI.</li>
-               <li class="list">Sample</li>
-                 <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                      <li class="list">Hv1a-his 3 7/18 (EP)</li>
-                      <li class="list">Sf1a-his 1 7/8 (EP)</li>
-                      <li class="list">OAIP-his 2 7/18 (EP)</li>
-                      <li class="list">Hv1a-lectin-his 4 7/8 (ES)</li>
-                      <li class="list">Sf1a-lectin-his 6 7/18 (ES)</li>
-                      <li class="list">OAIP-lectin-his 5 7/18(ES)</li>
-                      <li class="list">pSB1A3 (EP)	</li>
-                      <li class="list">pSB1A3 (ES)	</li>
-                 </ul>
-        </ul>
-        <p class="note-title">Ligation</p>
-           <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-               <li class="list">Ligase DNA fragment(7/23) into the pSB1A3</li>
-               <li class="list">Sample</li>
-                 <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                      <li class="list">Hv1a-his 3 7/18 (EP)</li>
-                      <li class="list">Sf1a-his 1 7/8 (EP)</li>
-                      <li class="list">OAIP-his 2 7/18 (EP)</li>
-                      <li class="list">Hv1a-lectin-his 4 7/8 (ES)</li>
-                      <li class="list">Sf1a-lectin-his 6 7/18 (ES)</li>
-                      <li class="list">OAIP-lectin-his 5 7/18(ES)</li>
-                      <li class="list">pSB1A3 (EP)	</li>
-                      <li class="list">pSB1A3 (ES)	</li>
-                 </ul>
-        </ul>
-       <p class="note-title">Transformation</p>
-           <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-               <li class="list">Transformation of ligated product.</li>
-               <li class="list">Sample</li>
-                 <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                      <li class="list">Hv1a-lectin-his A<sup>r</sup> 7/22</li>
-                      <li class="list">Sf1a-lectin-his A<sup>r</sup> 7/22</li>
-                      <li class="list">OAIP-lectin-his A<sup>r</sup> 7/22</li>
                   </ul>
          </ul>
@@ Line 1,611: / Line 956: @@
       <article id="July-24" class="note-item">
          <h2>July 24</h2><hr>
-        <p class="note-title">PCR</p>
+        <p class="note-title">Cloning</p>
             <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                <li class="list">PCR of ligated products(7/23) to amplify the insert gene.</li>
+                <li class="list">Cultivation</li>
+               <li class="list">Miniprep (Purify Plasmid)</li>
                 <li class="list">Sample</li>
                   <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                       <li class="list">Sf1a-lectin-his A<sup>r</sup> 7/23 1~4</li>
+                       <li class="list">Leucocyclicin Q + pTXB1</li>
-                      <li class="list">OAIP-lectin-his A<sup>r</sup> 7/23 5~8</li>
-                      <li class="list">Hv1a-lectin-his A<sup>r</sup> 7/23 9~12</li>
-                 </ul>
-        </ul>
-        <p class="note-title">Electrophoresis</p>
-           <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-               <li class="list">Electrophoresis of PCR products</li>
-               <li class="list">Sample</li>
-                 <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                      <li class="list">Sf1a-lectin-his A<sup>r</sup> 7/3 1~4</li>
-                      <li class="list">OAIP-lectin-his A<sup>r</sup> 7/23 5~8</li>
-                      <li class="list">Hv1a-lectin-his A<sup>r</sup> 7/23 9~12</li>
-                 </ul>
-        </ul>
-        <p class="note-title">Cultivation</p>
-           <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-               <li class="list">Cultivation of ligated product.(7/24)</li>
-               <li class="list">Sample</li>
-                 <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                      <li class="list">Sf1a-lectin-his A<sup>r</sup> 7/24 2.3</li>
-                      <li class="list">OAIP-lectin-his A<sup>r</sup> 7/24 5.7</li>
-                      <li class="list">Hv1a-lectin-his A<sup>r</sup> 7/24 9.12</li>
                   </ul>
          </ul>
@@ Line 1,648: / Line 969: @@
       <article id="July-25" class="note-item">
          <h2>July 25</h2><hr>
-        <p class="note-title">Miniprep</p>
+        <p class="note-title">Cloning</p>
             <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                <li class="list">Purify plasmid which have insert gene.(7/24)</li>
+                <li class="list">Sequencing plasmid</li>
                 <li class="list">Sample</li>
                   <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                       <li class="list">Sf1a-lectin-his A<sup>r</sup> 7/24 2.3</li>
+                       <li class="list">Leucocyclicin Q + pTXB1</li>
-                      <li class="list">OAIP-lectin-his A<sup>r</sup> 7/24 5.7</li>
-                      <li class="list">Hv1a-lectin-his A<sup>r</sup> 7/24 9.12</li>
-                 </ul>
-        </ul>
-        <p class="note-title">Digestion</p>
-           <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-               <li class="list">Digest the plasmid (7/25) with EcoRI and Spel to check whether the mini have insert gene fragment.</li>
-               <li class="list">Sample</li>
-                 <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                      <li class="list">Sf1a-lectin-his A<sup>r</sup>  2.3(ES)</li>
-                      <li class="list">OAIP-lectin-his A<sup>r</sup>  5.7(ES)</li>
-                      <li class="list">Hv1a-lectin-his A<sup>r</sup>  9.12(ES)</li>
-                 </ul>
-        </ul>
-        <p class="note-title">Electrophoresis</p>
-           <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-               <li class="list">Electrophoresis of ligated product.</li>
-               <li class="list">Sample</li>
-                 <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                      <li class="list">Sf1a-lectin-his A<sup>r</sup>  2.3(ES)</li>
-                      <li class="list">OAIP-lectin-his A<sup>r</sup>  5.7(ES)</li>
-                      <li class="list">Hv1a-lectin-his A<sup>r</sup>  9.12(ES)</li>
-                 </ul>
-        </ul>
-       <p class="note-title">Sequencing</p>
-           <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-               <li class="list">determine insert gene’s Sequence.</li>
-               <li class="list">Sample</li>
-                 <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                      <li class="list">Hv1a-lectin-his (The result is correct)</li>
-                      <li class="list">Hv1a-lectin-his (The result is correct)</li>
-                      <li class="list">OAIP - lectin-his (The result is correct)</li>
                   </ul>
          </ul>
@@ Line 1,696: / Line 981: @@
       <article id="July-26" class="note-item">
          <h2>July 26</h2><hr>
-        <p class="note-title">Prepare of CaCl<sub>2</sub></p>
+        <p class="note-title">None</p>
-           <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-               <li class="list">for competent cell</li>
-           </ul>
-        <p class="note-title">Rosetta-gami competent cell prepare</p>
-           <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-               <li class="list">for expression of disulfide bond.</li>
-           </ul>
-        <p class="note-title">Transformation</p>
-           <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-               <li class="list">Transformation of backbone plasmid into <i>BL21-rosetta-gami</i> check whether the competent cell is working.</li>
-               <li class="list">we choose pSB1A3 to transformation because <i>BL21-rosetta-gami</i> has C<sup>r</sup> and K<sup>r</sup>.</li>
-               <li class="list">Only write colonies. NO RFP EXPRESSION</li>
-           </ul>
       </article>
       <article id="July-27" class="note-item">
          <h2>July 27</h2><hr>
-         <p class="note-title">Transformation</p>
+         <p class="note-title">None</p>
-           <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-               <li class="list">Transformation of backbone plasmid into <i>BL21-rosetta-gami</i> check whether the competent cell is working.</li>
-               <li class="list">we choose pSB1A3 to transformation because <i>BL21-rosetta-gami</i> has C<sup>r</sup> and K<sup>r</sup>.</li>
-               <li class="list">Only write colonies. NO RFP EXPRESSION</li>
-           </ul>
-       <p class="note-title">Transformation</p>
-           <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-               <li class="list">Transformation of plasmid which have correct sequence to DH5α.</li>
-               <li class="list">Sample</li>
-                 <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                      <li class="list">Sf1a -his</li>
-                      <li class="list">OAIP-his</li>
-                      <li class="list">Hv1a -his</li>
-                 </ul>
-        </ul>
       </article>
       <article id="July-28" class="note-item">
          <h2>July 28</h2><hr>
-         <p class="note-title">Cultivation</p>
+         <p class="note-title">None</p>
-           <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-               <li class="list">Cultivation of correct colonies. (7/27)</li>
-                    <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                        <li class="list">Sf1a -his A<sup>r</sup></li>
-                        <li class="list">OAIP-his A<sup>r</sup></li>
-                        <li class="list">Hv1a -his A<sup>r</sup></li>
-                    </ul>
-           </ul>
-        <p class="note-title">Test competent cell</p>
-           <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-               <li class="list"><i>BL21-rosetta-gami</i> aren’t express RFP but write colonies. Therefore, we test competent cell whether they could save on A<sup>r</sup> agar plate.</li>
-               <li class="list">After cultivation about 14 hours, there are many write colonies. We think them may be contamination during preparing competent cell.</li>
-           </ul>
       </article>
       <article id="July-29" class="note-item">
          <h2>July 29</h2><hr>
-        <p class="note-title">Miniprep</p>
+        <p class="note-title">Expression</p>
             <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                <li class="list">Purify plasmid which have correct insert gene(7/28) to conserve and transform to <i>BL21-Rosetta-gami</i>.</li>
+                <li class="list">Transformation (Transform correct plasmid into <i>E. coli</i> ER2566)</li>
                      <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                         <li class="list">Sf1a -his A<sup>r</sup></li>
+                         <li class="list">Leucocyclicin Q + pTXB1</li>
-                        <li class="list">OAIP-his A<sup>r</sup></li>
-                        <li class="list">Hv1a -his A<sup>r</sup></li>
                      </ul>
             </ul>
-       <p class="note-title">Transformation</p>
-           <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-               <li class="list">Transformation of plasmid which have correct sequence to DH5α.</li>
-               <li class="list">Sample</li>
-                 <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                      <li class="list">Sf1a-lectin-his 7/24 A<sup>r</sup></li>
-                      <li class="list">OAIP-lectin-his 7/24 A<sup>r</sup></li>
-                      <li class="list">Hv1a-lectin-his 7/24 A<sup>r</sup></li>
-                 </ul>
-        </ul>
       </article>
       <article id="July-30" class="note-item">
          <h2>July 30</h2><hr>
-        <p class="note-title">Cultivation</p>
+        <p class="note-title">None</p>
-           <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-               <li class="list">Cultivation of correct colonies. (7/29)</li>
-                    <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                        <li class="list">Sf1a-lectin-his 7/29 A<sup>r</sup></li>
-                        <li class="list">OAIP-lectin-his 7/29 A<sup>r</sup></li>
-                        <li class="list">Hv1a-lectin-his 7/29 A<sup>r</sup></li>
-                    </ul>
-           </ul>
-       <p class="note-title">Miniprep</p>
-           <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-               <li class="list">Purify plasmid which have correct insert gene(7/30) to conserve and transform to  BL21-Rosetta-gami.</li>
-               <li class="list">Sample</li>
-                 <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                        <li class="list">Sf1a-lectin-his 7/29 A<sup>r</sup></li>
-                        <li class="list">OAIP-lectin-his 7/29 A<sup>r</sup></li>
-                        <li class="list">Hv1a-lectin-his 7/29 A<sup>r</sup></li>
-                 </ul>
-        </ul>
-       <p class="note-title">Preparation of Competent cell</p>
-           <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-               <li class="list">Rosetta-gami competent cell:</li>
-               <li class="list">Prepare the bacteria that can make disulfide bond</li>
-        </ul>
       </article>
       <article id="July-31" class="note-item">
@@ Line 1,819: / Line 1,018: @@
       <article id="August-1" class="note-item">
          <h2>August 1</h2><hr>
-        <p class="note-title">Transformation</p>
+        <p class="note-title">None</p>
-           <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-               <li class="list">Transformation of pSB1A3 to test the competent cell work.</li>
-               <li class="list">sample</li>
-                    <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                        <li class="list">PSB1A3</li>
-                    </ul>
-           </ul>
-       <p class="note-title">Transformation</p>
-           <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-               <li class="list">Transformation of plasmid which have correct sequence to <i>BL21-Rosetta-gami</i>.</li>
-               <li class="list">Sample</li>
-                 <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                        <li class="list">Hv1a-his</li>
-                        <li class="list">Sf1a-his</li>
-                        <li class="list">OAIP-his</li>
-                        <li class="list">Hv1a-lectin-his</li>
-                        <li class="list">Sf1a-lectin-his</li>
-                        <li class="list">OAIP-lectin-his</li>
-                 </ul>
-        </ul>
       </article>
       <article id="August-2" class="note-item">
          <h2>August 2</h2><hr>
-        <p class="note-title">Transformation</p>
+        <p class="note-title">None</p>
-           <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-               <li class="list">Transformation of plasmid which have correct sequence to <i>BL21-Rosetta-gami</i>.</li>
-               <li class="list">Sample</li>
-                 <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                        <li class="list">Sf1a-his 7/28 A<sup>r</sup></li>
-                        <li class="list">OAIP-his 7/28 A<sup>r</sup></li>
-                        <li class="list">Hv1a-lectin-his 7/29 A<sup>r</sup></li>
-                        <li class="list">Sf1a-lectin-his 7/29 A<sup>r</sup></li>
-                 </ul>
-        </ul>
       </article>
       <article id="August-3" class="note-item">
          <h2>August 3</h2><hr>
-        <p class="note-title">Cultivation</p>
+        <p class="note-title">None</p>
-           <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-               <li class="list">Cultivation of <i>BL21-Rosetta-gam</i> colonies for IPTG induction.</li>
-               <li class="list">sample</li>
-                    <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                        <li class="list">Hv1a-his 8/1 A<sup>r</sup></li>
-                        <li class="list">OAIP-lectin-his 8/1 A<sup>r</sup></li>
-                    </ul>
-           </ul>
-       <p class="note-title">Cultivation</p>
-           <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-               <li class="list">Cultivation of correct colonies again for mini extraction.</li>
-               <li class="list">Sample</li>
-                 <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                        <li class="list">Sf1a-his 7/27 A<sup>r</sup></li>
-                        <li class="list">OAIP-his 7/27 A<sup>r</sup></li>
-                        <li class="list">Hv1a-lectin-his 7/24 A<sup>r</sup></li>
-                        <li class="list">Sf1a-lectin-his 7/24 A<sup>r</sup></li>
-                 </ul>
-        </ul>
       </article>
       <article id="August-4" class="note-item">
          <h2>August 4</h2><hr>
-        <p class="note-title">Miniprep</p>
+        <p class="note-title">None</p>
-           <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-               <li class="list">Purify plasmid which have correct insert gene(8/3) to conserve and transform to <i>BL21-Rosetta-gami</i>.</li>
-               <li class="list">sample</li>
-                    <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                        <li class="list">OAIP-his 8/3  A<sup>r</sup></li>
-                        <li class="list">Sf1a-lectin-his 8/3 A<sup>r</sup></li>
-                        <li class="list">Hv1a-lectin-his 8/3 A<sup>r</sup></li>
-                    </ul>
-           </ul>
-       <p class="note-title">Cultivation</p>
-           <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-               <li class="list">Cultivation of correct colonies again for mini extraction.</li>
-               <li class="list">Sample</li>
-                 <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                        <li class="list">Sf1a-his 7/27 A<sup>r</sup></li>
-                 </ul>
-        </ul>
-       <p class="note-title">Transformation</p>
-           <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-               <li class="list">Transformation of plasmid which have correct sequence to <i>BL21-Rosetta-gami</i>.</li>
-               <li class="list">Sample</li>
-                 <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                      <li class="list">OAIP-his 8/3 A<sup>r</sup></li>
-                      <li class="list">Sf1a-lectin-his 8/3 A<sup>r</sup></li>
-                      <li class="list">Hv1a-lectin-his 8/3 A<sup>r</sup></li>
-                 </ul>
-        </ul>
-       <p class="note-title">IPTG induction</p>
-           <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-               <li class="list">To find out the best condition for IPTG induction.</li>
-               <li class="list">Sample</li>
-                 <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                      <li class="list">Hv1a-his 8/3 A<sup>r</sup></li>
-                      <li class="list">OAIP-lectin-his 8/3 A<sup>r</sup></li>
-                 </ul>
-               <li class="list">Condition<br>0μM, 250μM, 500μM, 750μM, 1000μM</li>
-               <li class="list">Add IPTG concentration and test OD<sub>600</sub><br>0μM, 250μM, 500μM, 750μM, 1000μM</li>
-        </ul>
       </article>
       <article id="August-5" class="note-item">
          <h2>August 5</h2><hr>
-        <p class="note-title">SDS-PAGE</p>
+        <p class="note-title">Expression</p>
             <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                <li class="list">Check the protein production after IPTG condition.</li>
+                <li class="list">Transformation (Transform correct plasmid into <i>E. coli</i> ER2566)</li>
                 <li class="list">sample</li>
                      <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                         <li class="list">Marker</li>
+                         <li class="list">Leucocyclicin Q + pTXB1</li>
-                        <li class="list">OAIP-lectin-his 250μM 2hr</li>
-                        <li class="list">OAIP-lectin-his 500μM 2hr</li>
-                        <li class="list">OAIP-lectin-his 750μM 2hr</li>
-                        <li class="list">OAIP-lectin-his 1000μM 2hr</li>
-                        <li class="list">OAIP-lectin-his 0μM 4hr</li>
-                        <li class="list">OAIP-lectin-his 250μM 4hr</li>
-                        <li class="list">OAIP-lectin-his 500μM 4hr</li>
-                        <li class="list">OAIP-lectin-his 750μM 4hr</li>
-                        <li class="list">OAIP-lectin-his 1000μM 4hr</li>
-                        <li class="list">Marker</li>
-                        <li class="list">OAIP-lectin-his 250μM 6hr</li>
-                        <li class="list">OAIP-lectin-his 500μM 6hr</li>
-                        <li class="list">OAIP-lectin-his 750μM 6hr</li>
-                        <li class="list">OAIP-lectin-his 1000μM 6hr</li>
-                        <li class="list">OAIP-lectin-his 0μM 8hr</li>
-                        <li class="list">OAIP-lectin-his 250μM 8hr</li>
-                        <li class="list">OAIP-lectin-his 500μM 8hr</li>
-                        <li class="list">OAIP-lectin-his 750μM 8hr</li>
-                        <li class="list">OAIP-lectin-his 1000μM 8hr</li>
                      </ul>
             </ul>
@@ Line 1,967: / Line 1,051: @@
       <article id="August-6" class="note-item">
          <h2>August 6</h2><hr>
-         <p class="note-title">None</p>
+         <p class="note-title">Expression</p>
+           <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
+               <li class="list">Cultivation (Cultivation of <i>E. coli</i> ER2566 colonies for IPTG induction)</li>
+               <li class="list">IPTG Induction (To test the O.D. levels of <i>E. coli</i> ER2566 to induce)Condition: O.D. = 0.5, 1.3</li>
+               <li class="list">sample</li>
+                    <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
+                        <li class="list">Leucocyclicin Q + pTXB1</li>
+                    </ul>
+           </ul>
       </article>
       <article id="August-7" class="note-item">
          <h2>August 7</h2><hr>
-         <p class="note-title">None</p>
+         <p class="note-title">Expression</p>
+           <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
+               <li class="list">SDS-PAGE (To check the protein production after induction)</li>
+               <li class="list">sample</li>
+                    <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
+                        <li class="list">Leucocyclicin Q + pTXB1</li>
+                    </ul>
+           </ul>
       </article>
       <article id="August-8" class="note-item">
@@ Line 1,987: / Line 1,086: @@
       <article id="August-11" class="note-item">
          <h2>August 11</h2><hr>
-         <p class="note-title">None</p>
+         <p class="note-title">Cloning</p>
+           <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
+               <li class="list">Amplify the insert gene (Pfu PCR)</li>
+               <li class="list">Electrophoresis (To check PCR products)</li>
+               <li class="list">sample</li>
+                    <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
+                        <li class="list">Enterocin B</li>
+                        <li class="list">Enterocin 96 </li>
+                        <li class="list">Durancin TW49M </li>
+                    </ul>
+           </ul>
       </article>
       <article id="August-12" class="note-item">
          <h2>August 12</h2><hr>
-         <p class="note-title">None</p>
+         <p class="note-title">Cloning</p>
+           <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
+               <li class="list">Digestion of PCR products (Including insert and backbone pTXB1)</li>
+               <li class="list">Ligation of digestion products (Ligase the DNA fragment into backbone pTXB1)</li>
+               <li class="list">Transformation of ligation products (Transform into <i>E. coli</i> DH5α)</li>
+               <li class="list">sample</li>
+                    <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
+                        <li class="list">Enterocin B + pTXB1</li>
+                        <li class="list">Enterocin 96  + pTXB1</li>
+                        <li class="list">Durancin TW-49M  + pTXB1</li>
+                    </ul>
+           </ul>
+        <p class="note-title">Growth curve</p>
+           <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
+               <li class="list">Observe the growth of <i>Bacillus subtilis</i> in different temperature, pH, salinityCondition:</li>
+               <li class="list">1.Temp: 37°C , pH:7 , salinity: 0.17M</li>
+               <li class="list">2.Temp: 30°C , pH:9 , salinity: 0.5M</li>
+               <li class="list">3.Temp: 25°C , pH:5 , salinity: 0.25M</li>
+           </ul>
       </article>
       <article id="August-13" class="note-item">
          <h2>August 13</h2><hr>
-         <p class="note-title">None</p>
+         <p class="note-title">Cloning</p>
+           <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
+               <li class="list">Taq PCR (PCR of ligation products to amplify insert gene)</li>
+               <li class="list">Electrophoresis (To check PCR products)</li>
+               <li class="list">sample</li>
+                    <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
+                        <li class="list">Enterocin B + pTXB1</li>
+                        <li class="list">Enterocin 96  + pTXB1</li>
+                        <li class="list">Durancin TW-49M  + pTXB1</li>
+                    </ul>
+           </ul>
       </article>
@@ Line 2,001: / Line 1,138: @@
       <article id="August-14" class="note-item">
          <h2>August 14</h2><hr>
-        <p class="note-title">Transformation</p>
+        <p class="note-title">Cloning</p>
             <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                <li class="list">Transformation of plasmid which have correct sequence to <i>BL21-Rosetta-gami</i>.</li>
+                <li class="list">Cultivation</li>
+               <li class="list">Miniprep (Purify Plasmid)</li>
                 <li class="list">Sample</li>
                   <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                      <li class="list">Sf1a-his 7/29  A<sup>r</sup></li>
+                        <li class="list">Enterocin B + pTXB1</li>
-                      <li class="list">OAIP-his 8/4 A<sup>r</sup></li>
+                        <li class="list">Enterocin 96  + pTXB1</li>
-                      <li class="list">Hv1a-his 7/29  A<sup>r</sup></li>
+                        <li class="list">Durancin TW-49M  + pTXB1</li>
-                      <li class="list">Sf1a-lectin-his 8/4  A<sup>r</sup></li>
-                      <li class="list">OAIP-lectin-his 7/30  A<sup>r</sup></li>
-                      <li class="list">Hv1a-lectin-his 8/4  A<sup>r</sup></li>
                   </ul>
-        </ul>
+           </ul>
+        <p class="note-title">Expression</p>
-       <p class="note-title">PCR</p>
             <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                <li class="list">PCR of transformation colonies to check the base pair.</li>
+                <li class="list">Transformation (Transform correct plasmid into <i>E. coli</i> ER2566)to test different cultivation temperature</li>
-               <li class="list">Sample</li>
-                 <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                      <li class="list">Hv1a-his A<sup>r</sup> 1~6</li>
-                      <li class="list">Hv1a-lectin-his A<sup>r</sup> 1~6</li>
-                      <li class="list">Sf1a-lectin-his A<sup>r</sup> 1~6</li>
-                      <li class="list">OAIP-lectin-his A<sup>r</sup> 1~6</li>
-                 </ul>
          </ul>
       </article>
@@ Line 2,030: / Line 1,157: @@
       <article id="August-15" class="note-item">
          <h2>August 15</h2><hr>
-        <p class="note-title">PCR</p>
+        <p class="note-title">Cloning</p>
             <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                <li class="list">PCR of ligated products to amplify the insert gene.</li>
+                <li class="list">Sequencing plasmid</li>
                 <li class="list">Sample</li>
                   <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                      <li class="list">Hv1a-his 8/14  A<sup>r</sup> 1~6</li>
+                        <li class="list">Enterocin B + pTXB1</li>
-                      <li class="list">Hv1a-lectin-his 8/14 A<sup>r</sup> 1~6</li>
+                        <li class="list">Enterocin 96  + pTXB1</li>
-                      <li class="list">Sf1a- lectin-his 8/14  A<sup>r</sup> 1~6</li>
+                        <li class="list">Durancin TW-49M  + pTXB1</li>
-                      <li class="list">Sf1a-lectin-his 8/4  A<sup>r</sup> 1~6</li>
-                      <li class="list">OAIP-lectin-his 7/30  A<sup>r</sup> 1~6</li>
-                      <li class="list">OAIP- lectin-his 8/14  A<sup>r</sup> 1~6</li>
                   </ul>
          </ul>
-        <p class="note-title">Electrophoresis</p>
+        <p class="note-title">Expression</p>
             <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                <li class="list">Electrophoresis of PCR products to check the insert gene base pair</li>
+                <li class="list">Cultivation (Cultivation of <i>E. coli</i> ER2566 colonies for IPTG induction)</li>
-               <li class="list">Sample</li>
+                <li class="list">IPTG Induction (To test the temperature of cultivation after induction)Condition: temp = 37, 30, 13.5°C</li>
-                 <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                      <li class="list">Hv1a-his 8/15 A<sup>r</sup> 1~6</li>
-                      <li class="list">Hv1a-lectin-his 8/15 A<sup>r</sup> 1~6</li>
-                      <li class="list">Sf1a- lectin-his 8/15 A<sup>r</sup> 1~6</li>
-                      <li class="list">OAIP- lectin-his 8/15 A<sup>r</sup> 1~6</li>
-                 </ul>
-        </ul>
-       <p class="note-title">Transformation</p>
-           <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-               <li class="list">Transformation of plasmid which have correct sequence to <i>BL21-Rosetta-gami</i>.</li>
-                <li class="list">Sample</li>
-                 <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                      <li class="list">Sf1a-his 7/29 A<sup>r</sup></li>
-                      <li class="list">OAIP-his 8/4 A<sup>r</sup></li>
-                 </ul>
-        </ul>
-       <p class="note-title">PCR</p>
-           <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-               <li class="list">PCR of ligated products to amplify the insert gene.</li>
-               <li class="list">Sample</li>
-                 <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                      <li class="list">Sf1a- lectin-his 8/15 A<sup>r</sup> 1~6</li>
-                      <li class="list">OAIP- lectin-his 8/15 A<sup>r</sup> 1~6</li>
-                 </ul>
-        </ul>
-       <p class="note-title">Electrophoresis</p>
-           <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-               <li class="list">Electrophoresis of PCR products to check the insert gene base pair.</li>
-               <li class="list">Sample</li>
-                 <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                      <li class="list">Sf1a- lectin-his 8/15 A<sup>r</sup> 1~6</li>
-                      <li class="list">OAIP- lectin-his 8/15 A<sup>r</sup> 1~6</li>
-                 </ul>
-        </ul>
-       <p class="note-title">SDS-PAGE</p>
-           <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-               <li class="list">Check the production of IPTG induction (8/4) again.</li>
-               <li class="list">No target protein expression.</li>
          </ul>
       </article>
@@ Line 2,095: / Line 1,177: @@
       <article id="August-16" class="note-item">
          <h2>August 16</h2><hr>
-        <p class="note-title">Cultivation</p>
+        <p class="note-title">Expression</p>
             <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                <li class="list">Cultivation of <i>BL21-Rosetta-gami</i> colonies for IPTG induction.</li>
+                <li class="list">SDS-PAGE (To check the protein production after induction)Check different temperature</li>
-               <li class="list">Sample</li>
+                <li class="list">IPTG Induction (To test the concentration of IPTG)Condition: concentration = 200, 400, 600, 800, 1000μM</li>
-                 <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                      <li class="list">Hv1a-his 8/15  A<sup>r</sup></li>
-                      <li class="list">Hv1a-lectin-his 8/15 A<sup>r</sup></li>
-                      <li class="list">Sf1a- lectin-his 8/15  A<sup>r</sup></li>
-                      <li class="list">OAIP-lectin-his 8/15  A<sup>r</sup></li>
-                 </ul>
-        </ul>
-       <p class="note-title">IPTG induction</p>
-           <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-               <li class="list">To check the protein expression for IPTG induction.</li>
-                <li class="list">Sample</li>
-                 <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                      <li class="list">Hv1a-his 8/16  A<sup>r</sup></li>
-                      <li class="list">Hv1a-lectin-his 8/16 A<sup>r</sup></li>
-                      <li class="list">Sf1a- lectin-his 8/16  A<sup>r</sup></li>
-                      <li class="list">OAIP-lectin-his 8/16 A<sup>r</sup></li>
-                 </ul>
-               <li class="list">Condition<br>0μM, 500μM</li>
-               <li class="list">Add IPTG 500μM and test OD<sub>600</sub> after 4hr.</li>
-        </ul>
-       <p class="note-title">Sonication and SDS-PAGE</p>
-           <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-               <li class="list">Sonication and run SDS-PAGE to check peptides expression level after the IPTG induction.</li>
-               <li class="list">Sample(8/16)</li>
-                 <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                      <li class="list">Hv1a-his 0μM 2hr</li>
-                      <li class="list">Hv1a-his 500μM 2hr</li>
-                      <li class="list">Sf1a-lectin-his 0μM 4hr</li>
-                      <li class="list">Sf1a-lectin-his 500μM 4hr</li>
-                      <li class="list">Hv1a-lectin-his 0μM 2hr</li>
-                      <li class="list">Hv1a-lectin-his 500μM 2hr</li>
-                      <li class="list">OAIP-lectin-his 0μM 4hr</li>
-                      <li class="list">OAIP-lectin-his 500μM 4hr</li>
-                 </ul>
-               <li class="list">Failed. Do it again</li>
-               <li class="list">Sample</li>
-                 <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                      <li class="list">Hv1a-his 0μM 2hr</li>
-                      <li class="list">Hv1a-his 500μM 2hr</li>
-                      <li class="list">Sf1a-lectin-his 0μM 4hr</li>
-                      <li class="list">Sf1a-lectin-his 500μM 4hr</li>
-                      <li class="list">Hv1a-lectin-his 0μM 2hr</li>
-                      <li class="list">Hv1a-lectin-his 500μM 2hr</li>
-                      <li class="list">OAIP-lectin-his 0μM 4hr</li>
-                      <li class="list">OAIP-lectin-his 500μM 4hr</li>
-                 </ul>
          </ul>
       </article>
@@ Line 2,152: / Line 1,186: @@
       <article id="August-17" class="note-item">
          <h2>August 17</h2><hr>
-            <p class="note-title">IPTG induction</p>
+            <p class="note-title">Expression</p>
             <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                <li class="list">To check the protein expression for IPTG induction.</li>
+                <li class="list">SDS-PAGE (To check the protein production after induction) -> Check different IPTG concentration</li>
+           </ul>
+     </article>
+     <article id="August-18" class="note-item">
+        <h2>August 18</h2><hr>
+       <p class="note-title">Transformation</p>
+           <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
+               <li class="list">Transforming backbone pTXB1 into <i>E. coli</i> ER2566 to produce protein as our negative control in verifying the function of our target peptide.</li>
+               <li class="list">Transforming backbone pTXB1 containing bacteriocin gene into <i>E. coli</i> ER2566 to produce our target protein as a bio-stimulator.</li>
                 <li class="list">Sample</li>
                   <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                       <li class="list">Hv1a-his 8/17 A<sup>r</sup></li>
+                       <li class="list">backbone pTXB1 mini</li>
-                       <li class="list">Hv1a-lectin-his </li>
+                       <li class="list">backbone pTXB1 containing Enterocin B gene mini</li>
-                       <li class="list">Sf1a- lectin-his</li>
+                       <li class="list">backbone pTXB1 containing Enterocin 96 gene mini</li>
-                       <li class="list">OAIP- lectin-his</li>
+                       <li class="list">backbone pTXB1 containing Leucocyclicin Q gene mini</li>
                   </ul>
-               <li class="list">Condition<br>
+         <p class="note-title">Cultivation</p>
-μM, 500μM
-</li>
-               <li class="list">Add IPTG 500μM and test OD<sub>600</sub> after 4hr.</li>
-         </ul>
-       <p class="note-title">Sonication and SDS-PAGE</p>
             <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                <li class="list">Sonication and run SDS-PAGE to check peptides expression level after the IPTG induction.</li>
+                <li class="list">Cultivate the <i>E. coli</i> ER2566 at 37°C cultivating <i>Bacillus subtilis</i> ER2566 at 37°C carrying the backbone pTXB1 to produce protein as our negative control in verifying the function of our target peptide.</li>
+               <li class="list">Cultivate the <i>E. coli</i> ER2566 at 37°C cultivating <i>E. coli</i> ER2566 at 37°C carrying the backbone pTXB1 containing bacteriocin gene to produce our target protein as a bio-stimulator.</li>
                 <li class="list">Sample</li>
                   <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                       <li class="list">Hv1a-his 0μM 2hr</li>
+                       <li class="list">Fresh colony of <i>E. coli</i> ER2566 carrying the backbone pTXB1 from an overnight plate</li>
-                      <li class="list">Hv1a-his 500μM 2hr</li>
+                       <li class="list">Fresh colony of <i>E. coli</i> ER2566 carrying the backbone pTXB1 containing Enterocin B gene from an overnight plate</li>
-                       <li class="list">Sf1a-lectin-his 0μM 4hr</li>
+                       <li class="list">Fresh colony of <i>E. coli</i> ER2566 carrying the backbone pTXB1 containing Enterocin 96 gene from an overnight plate</li>
-                      <li class="list">Sf1a-lectin-his 500μM 4hr</li>
+                       <li class="list">Fresh colony of <i>E. coli</i> ER2566 carrying the backbone pTXB1 containing Leucocyclicin Q gene from an overnight plate</li>
-                       <li class="list">Hv1a-lectin-his 0μM 2hr</li>
+                  </ul>
-                      <li class="list">Hv1a-lectin-his 500μM 2hr</li>
+         <p class="note-title">IPTG Induction</p>
-                       <li class="list">OAIP-lectin-his 0μM 4hr</li>
-                      <li class="list">OAIP-lectin-his 500μM 4hr</li>
-                  </ul>
-         </ul>
-     </article>
-     <article id="August-18" class="note-item">
-        <h2>August 18</h2><hr>
-       <p class="note-title">SDS-PAGE</p>
             <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                <li class="list">To Check protein disulfide bond folding.</li>
+                <li class="list">Inducting <i>E. coli</i> ER2566 carrying the backbone pTXB1 after cultivating at 37°C to produce protein as our negative control in verifying the function of our target peptide</li>
-                <li class="list">β-me make disulfide bond not folding.</li>
+                <li class="list">Inducting <i>E. coli</i> ER2566 carrying the backbone pTXB1 containing bacteriocin gene after cultivating at 37°C to produce our target protein as a bio-stimulator.</li>
                 <li class="list">Sample</li>
                   <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                       <li class="list">Hv1a-lectin-his 0μM + β-me</li>
+                       <li class="list">Culture of <i>E. coli</i> ER2566 carrying the backbone pTXB1</li>
-                      <li class="list">Hv1a-lectin-his 500μM + β-me</li>
+                       <li class="list">Culture of <i>E. coli</i> ER2566 carrying the backbone pTXB1 containing Enterocin B gene</li>
-                       <li class="list">Hv1a-lectin-his 500μM</li>
+                       <li class="list">Culture of <i>E. coli</i> ER2566 carrying the backbone pTXB1 containing Enterocin 96 gene</li>
-                      <li class="list">Sf1a-lectin-his 0μM + β-me</li>
+                       <li class="list">Culture of <i>E. coli</i> ER2566 carrying the backbone pTXB1 containing Leucocyclicin Q gene</li>
-                       <li class="list">Sf1a-lectin-his 500μM + β-me</li>
+                  </ul>
-                      <li class="list">Sf1a-lectin-his 500μM</li>
-                       <li class="list">OAIP-lectin-his 0μM + β-me</li>
-                      <li class="list">OAIP-lectin-his 500μM + β-me</li>
-                      <li class="list">OAIP-lectin-his 500μM</li>
-                  </ul>
-        </ul>
       </article>
       <article id="August-19" class="note-item">
          <h2>August 19</h2><hr>
-         <p class="note-title">SDS-PAGE</p>
+         <p class="note-title">Cloning</p>
             <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                <li class="list">To Check protein disulfide bond folding with the induction of control, Rosetta-gami stain.</li>
+                <li class="list">Digestion of PCR products (Including insert and backbone pET30a)</li>
-                <li class="list">β-me make disulfide bond not folding.</li>
+                <li class="list">Ligation of digestion products (Ligase the DNA fragment into backbone pET30a)</li>
                 <li class="list">Sample(8/17)</li>
                   <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                       <li class="list">Hv1a-lectin-his 0μM + β-me</li>
+                       <li class="list">Enterocin B + pET30a</li>
-                       <li class="list">Hv1a-lectin-his 500μM + β-me</li>
+                       <li class="list">Enterocin 96 + pET30a</li>
-                       <li class="list">Hv1a-lectin-his 500μM</li>
+                       <li class="list">Bovicin HJ50 + pET30a</li>
-                      <li class="list">Sf1a-lectin-his 0μM + β-me</li>
+                       <li class="list">Durancin TW-49M + pET30a</li>
-                       <li class="list">Sf1a-lectin-his 500μM + β-me</li>
+                       <li class="list">Lacticin Z + pET30a</li>
-                       <li class="list">Sf1a-lectin-his 500μM</li>
+                       <li class="list">Leucocyclicin Q + pET30a</li>
-                      <li class="list">Rosetta-gami + β-me</li>
-                       <li class="list">OAIP-lectin-his 0μM + β-me</li>
-                      <li class="list">OAIP-lectin-his 500μM + β-me</li>
-                      <li class="list">OAIP-lectin-his 500μM</li>
                   </ul>
          </ul>
-     </article>
+         <p class="note-title">Sonication</p>
-     <article id="August-20" class="note-item">
-         <h2>August 20</h2><hr>
-       <p class="note-title">Sonication and SDS-PAGE</p>
             <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                <li class="list">Sonication and run SDS-PAGE to check peptides expression level after the IPTG induction.</li>
+                <li class="list">Breaking <i>E. coli</i> ER2566 carrying the backbone pTXB1 and pTXB1 containing bacteriocin gene after cultivating to harvest the protein by sonication.</li>
                 <li class="list">Sample</li>
                   <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                       <li class="list">Hv1a-his A<sup>r</sup></li>
+                       <li class="list">Culture of <i>E. coli</i> ER2566 carrying the backbone pTXB1 induced by IPTG</li>
-                       <li class="list">Hv1a-his A<sup>r</sup></li>
+                       <li class="list">Culture of <i>E. coli</i> ER2566 carrying the backbone pTXB1 containing Enterocin B gene induced by IPTG </li>
-                       <li class="list">Sf1a-lectin-his A<sup>r</sup></li>
+                       <li class="list">Culture of <i>E. coli</i> ER2566 carrying the backbone pTXB1 containing Enterocin 96 gene induced by IPTG </li>
-                       <li class="list">Sf1a-lectin-his A<sup>r</sup></li>
+                       <li class="list">Culture of <i>E. coli</i> ER2566 carrying the backbone pTXB1 containing Leucocyclicin Q gene induced by IPTG </li>
-                      <li class="list">Hv1a-lectin-his A<sup>r</sup></li>
-                      <li class="list">Hv1a-lectin-his A<sup>r</sup></li>
-                      <li class="list">OAIP-lectin-his A<sup>r</sup></li>
-                      <li class="list">OAIP-lectin-his A<sup>r</sup></li>
                   </ul>
          </ul>
+        <p class="note-title">Protein quantification & SDS-PAGE</p>
-       <p class="note-title">Cultivation</p>
             <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                <li class="list">Cultivation of sample BL21-Rosetta-gami colonies for protein purification.</li>
+                <li class="list">Quantificating the protein expressing the backbone pTXB1 and pTXB1 containing bacteriocin gene by Bradford method and running SDS-PAGE to check the protein expression.</li>
                 <li class="list">Sample</li>
                   <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                       <li class="list">Hv1a-his A<sup>r</sup> 8/20</li>
+                       <li class="list">Protein expressing the backbone pTXB1 induced by IPTG</li>
-                       <li class="list">Hv1a-lectin-his  A<sup>r</sup> 8/20</li>
+                       <li class="list">Protein expressing the backbone pTXB1 containing Enterocin B gene induced by IPTG </li>
-                       <li class="list">Sf1a-lectin-hiss A<sup>r</sup> 8/20</li>
+                       <li class="list">Protein expressing the backbone pTXB1 containing Enterocin 96 gene induced by IPTG </li>
-                       <li class="list">OAIP-lectin-his A<sup>r</sup> 8/20</li>
+                       <li class="list">Protein expressing the backbone pTXB1 containing Leucocyclicin Q gene induced by IPTG </li>
                   </ul>
          </ul>
+     </article>
-        <p class="note-title">SDS-PAGE</p>
+     <article id="August-20" class="note-item">
+        <h2>August 20</h2><hr>
+        <p class="note-title">Cloning</p>
             <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                <li class="list">To Check protein disulfide bond folding with the induction of control, <i>Rosetta-gami stain</i>.</li>
+                <li class="list">Transformation of ligation products (Transform into <i>E. coli</i> DH5α)</li>
+               <li class="list">Taq PCR (PCR of ligation products to amplify insert gene)</li>
+               <li class="list">Electrophoresis (To check PCR products)</li>
                 <li class="list">Sample</li>
                   <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                       <li class="list">Hv1a-his 8/19</li>
+                       <li class="list">Enterocin B + pET30a</li>
-                       <li class="list">Sf1a-his 8/19</li>
+                       <li class="list">Enterocin 96 + pET30a</li>
-                       <li class="list">OAIP-his 8/19</li>
+                      <li class="list">Bovicin HJ50 + pET30a</li>
+                       <li class="list">Durancin TW-49M + pET30a</li>
+                      <li class="list">Lacticin Z + pET30a</li>
+                      <li class="list">Leucocyclicin Q + pET30a</li>
                   </ul>
-</ul>
+        </ul>
       </article>
       <article id="August-21" class="note-item">
          <h2>August 21</h2><hr>
-        <p class="note-title">IPTG Induction and Sonication</p>
+        <p class="note-title">Cloning</p>
             <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                <li class="list">To check the protein expression for IPTG induction. (Second round)</li>
+                <li class="list">Cultivation</li>
+               <li class="list">Miniprep (Purify Plasmid)</li>
                 <li class="list">Sample</li>
                   <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                       <li class="list">Hv1a-his A<sup>r</sup> 8/20</li>
+                       <li class="list">Enterocin B + pET30a</li>
-                       <li class="list">Hv1a-lectin-his A<sup>r</sup> 8/20</li>
+                       <li class="list">Enterocin 96 + pET30a</li>
-                       <li class="list">Sf1a-his  A<sup>r</sup> 8/17</li>
+                       <li class="list">Bovicin HJ50 + pET30a</li>
-                       <li class="list">Sf1a-lectin-his A<sup>r</sup> 8/20</li>
+                       <li class="list">Durancin TW-49M + pET30a</li>
-                       <li class="list">OAIP-his A<sup>r</sup> 8/17</li>
+                       <li class="list">Lacticin Z + pET30a</li>
-                       <li class="list">OAIP-lectin-his A<sup>r</sup> 8/20</li>
+                       <li class="list">Leucocyclicin Q + pET30a</li>
                   </ul>
-               <li class="list">Condition<br>0μM, 500μM</li>
-               <li class="list">Add IPTG 500μM and test OD<sub>600</sub> after 4hr.</li>
-        </ul>
       </article>
       <article id="August-22" class="note-item">
          <h2>August 22</h2><hr>
-        <p class="note-title">Cultivation</p>
+        <p class="note-title">Cloning</p>
             <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                <li class="list">Cultivation of sample <i>BL21-Rosetta-gami</i> colonies for protein purification.</li>
+                <li class="list">Sequencing plasmid</li>
                 <li class="list">Sample</li>
                   <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                       <li class="list">	Hv1a-lectin-his 8/20  A<sup>r</sup></li>
+                       <li class="list">Enterocin B + pET30a</li>
-                 </ul>
+                       <li class="list">Enterocin 96 + pET30a</li>
-        </ul>
+                       <li class="list">Bovicin HJ50 + pET30a</li>
+                       <li class="list">Durancin TW-49M + pET30a</li>
-       <p class="note-title">Cultivation</p>
+                       <li class="list">Lacticin Z + pET30a</li>
-           <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
+                       <li class="list">Leucocyclicin Q + pET30a</li>
-               <li class="list">Cultivation of sample BL21-Rosetta-gami colonies for protein purification.</li>
-               <li class="list">Sample</li>
-                 <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                       <li class="list"><i>BL21-Rosetta-gami</i> competent cell</li>
-                       <li class="list">Cultivation 11 hours</li>
-                 </ul>
-        </ul>
-       <p class="note-title">IPTG Induction</p>
-           <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-               <li class="list">Prepare for the purify the protein from sample <i>BL21-Rosetta-gami</i> colonies.</li>
-               <li class="list">Sample</li>
-                 <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                       <li class="list">Sf1a-his Ar 8/20 A<sup>r</sup></li>
-                 </ul>
-               <li class="list">Condition<br>0μM, 1000μM</li>
-               <li class="list">Add IPTG 500μM and test OD<sub>600</sub> after 4hr.</li>
-        </ul>
-       <p class="note-title">Sonication</p>
-           <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-               <li class="list">Sample</li>
-                 <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                       <li class="list">Hv1a-lectin-his 8/22</li>
-                       <li class="list">BL21-Rosetta-gami 8/22</li>
-                      <li class="list">Sf1a-lectin-his 0μM 8/22</li>
-                      <li class="list">Sf1a-lectin-his 1000μM 8/22</li>
                   </ul>
          </ul>
@@ Line 2,338: / Line 1,324: @@
       <article id="August-23" class="note-item">
          <h2>August 23</h2><hr>
-        <p class="note-title">Protein purification(8/22-8/23)</p>
+        <p class="note-title">None</p>
-           <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-               <li class="list">Sample</li>
-                 <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                      <li class="list">Hv1a-lectin-his</li>
-                 </ul>
-               <li class="list">Collect：<br>pass through solution, wash through solution(W1~3), elution solution(E1~10), filtered NaCl solution, and filtered ddH<sub>2</sub>O solution.</li>
-        </ul>
-       <p class="note-title">Cultivation and IPTG induction</p>
-           <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-               <li class="list">Cultivation of sample <i>BL21-Rosetta-gami</i> colonies for protein purification.</li>
-               <li class="list">Sample</li>
-                 <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                      <li class="list">Hv1a-his A<sup>r</sup> 8/20</li>
-                      <li class="list">Sf1a-lectin-his A<sup>r</sup> 8/20</li>
-                 </ul>
-               <li class="list">Add 1000μM IPTG and test OD<sub>600</sub> </li>
-        </ul>
-       <p class="note-title">Sonication</p>
-           <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-               <li class="list">Sample</li>
-                 <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                      <li class="list">Hv1a-his A<sup>r</sup> 8/23</li>
-                      <li class="list">Sf1a-lectin-his A<sup>r</sup> 8/23</li>
-                 </ul>
-        </ul>
       </article>
       <article id="August-24" class="note-item">
          <h2>August 24</h2><hr>
-       <p class="note-title">Protein purification(8/23-8/24)</p>
+        <p class="note-title">Transformation</p>
             <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
+               <li class="list">Transforming backbone pTXB1 containing bacteriocin gene into <i>E. coli</i> ER2566 to produce our target protein as a bio-stimulator.</li>
                 <li class="list">Sample</li>
                   <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                       <li class="list">Hv1a-lectin-his</li>
+                       <li class="list">backbone pTXB1 containing Bovicin HJ50 gene mini</li>
+                      <li class="list">backbone pTXB1 containing Durancin TW-49M gene mini</li>
                   </ul>
-               <li class="list">Collect：<br>pass through solution, wash through solution(W1~3), elution solution(E1~10), filtered NaCl solution, and filtered ddH<sub>2</sub>O solution.</li>
+           </ul>
-         </ul>
+         <p class="note-title">Cultivation</p>
-       <p class="note-title">Cultivation and IPTG induction</p>
             <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                <li class="list">Cultivation of sample <i>BL21-Rosetta-gami</i> colonies for protein purification.</li>
+                <li class="list">Cultivate the <i>E. coli</i> ER2566 at 37°C cultivating <i>E. coli</i> ER2566 at 37°C carrying the backbone pTXB1 containing bacteriocin gene to produce our target protein as a bio-stimulator.</li>
                 <li class="list">Sample</li>
                   <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                       <li class="list">Sf1a-his A<sup>r</sup> 8/20</li>
+                       <li class="list">Fresh colony of <i>E. coli</i> ER2566 carrying the backbone pTXB1 containing Bovicin HJ50 gene from an overnight plate</li>
-                       <li class="list">Sf1a-lectin-his A<sup>r</sup> 8/20</li>
+                       <li class="list">Fresh colony of <i>E. coli</i> ER2566 carrying the backbone pTXB1 containing Durancin TW-49M gene from an overnight plate</li>
-                 </ul>
+                  </ul>
-               <li class="list">Refresh to approximately OD<sub>600</sub> 0.7</li>
+           </ul>
-               <li class="list">Add 1000μM IPTG and test OD<sub>600</sub></li>
+         <p class="note-title">IPTG Induction</p>
-               <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                      <li class="list">Sf1a-his: 0.966</li>
-                      <li class="list">Sf1a-lectin-his: 1.363</li>
-                  </ul>
-               <li class="list">Certification, and add 2mL binding buffer and 200μM PMSF for sonication. Repeat three times.</li>
-         </ul>
-       <p class="note-title">Sonication</p>
             <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
+               <li class="list">Inducting <i>E. coli</i> ER2566 carrying the backbone pTXB1 containing bacteriocin gene after cultivating at 37°C to produce our target protein as a bio-stimulator.</li>
                 <li class="list">Sample</li>
                   <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                       <li class="list">Sf1a-his A<sup>r</sup> 8/24</li>
+                       <li class="list">Culture of <i>E. coli</i> ER2566 carrying the backbone pTXB1 containing Bovicin HJ50 gene</li>
-                       <li class="list">Sf1a-lectin-his A<sup>r</sup> 8/24</li>
+                       <li class="list">Culture of <i>E. coli</i> ER2566 carrying the backbone pTXB1 containing Durancin TW-49M gene</li>
                   </ul>
+            </ul>
-        </ul>
-       <p class="note-title">SDS-PAGE</p>
-            <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-               <li class="list">Check the purification protein.</li>
-               <li class="list">Sample</li>
-               <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                      <li class="list">Hv1a-lectin-his(8/22-8/23)</li>
-               </ul>
-        </ul>
       </article>
       <article id="August-25" class="note-item">
          <h2>August 25</h2><hr>
-       <p class="note-title">Protein purification(8/24-8/25)</p>
+               <p class="note-title">Sonication</p>
             <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
+               <li class="list">Breaking <i>E. coli</i> ER2566 carrying the backbone pTXB1 containing bacteriocin gene after cultivating to harvest the protein by sonication.</li>
                 <li class="list">Sample</li>
                   <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                       <li class="list">Hv1a-lectin-his</li>
+                       <li class="list">Culture of <i>E. coli</i> ER2566 carrying the backbone pTXB1 containing Bovicin HJ50 gene induced by IPTG </li>
+                      <li class="list">Culture of <i>E. coli</i> ER2566 carrying the backbone pTXB1 containing Durancin TW-49M gene induced by IPTG </li>
                   </ul>
-               <li class="list">Collect：<br>pass through solution, wash through solution(W1~3), elution solution(E1~10), filtered NaCl solution, and filtered ddH<sub>2</sub>O solution.</li>
          </ul>
-       <p class="note-title">SDS-PAGE</p>
+        <p class="note-title">Protein quantification & SDS-PAGE</p>
             <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                <li class="list">Check the purification protein.</li>
+                <li class="list">Quantificating the protein expressing the backbone pTXB1 containing bacteriocin gene by Bradford method and running SDS-PAGE to check the protein expression.</li>
                 <li class="list">Sample</li>
-               <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
+                 <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                       <li class="list">Hv1a-lectin-his(8/23-8/24)</li>
+                       <li class="list">Protein expressing the backbone pTXB1 containing Bovicin HJ50 gene induced by IPTG </li>
-               </ul>
+                      <li class="list">Protein expressing the backbone pTXB1 containing Durancin TW-49M gene induced by IPTG </li>
+                 </ul>
          </ul>
       </article>
@@ Line 2,440: / Line 1,382: @@
       <article id="August-26" class="note-item">
          <h2>August 26</h2><hr>
-        <p class="note-title">Cultivation</p>
+        <p class="note-title">None</p>
-           <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-               <li class="list">Cultivation of sample <i>BL21-Rosetta-gami</i> colonies for protein purification.</li>
-               <li class="list">Sample</li>
-                 <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                      <li class="list">Hv1a-his A<sup>r</sup> 8/15</li>
-                      <li class="list">Hv1a-lectin-his A<sup>r</sup> 8/15</li>
-                      <li class="list">Sf1a-lectin-his A<sup>r</sup> 8/15</li>
-                      <li class="list">OAIP-lectin-his A<sup>r</sup> 8/15</li>
-                 </ul>
-        </ul>
-       <p class="note-title">Sonication</p>
-           <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-               <li class="list">Sample</li>
-                 <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                      <li class="list">Sf1a-his A<sup>r</sup> 8/24</li>
-                      <li class="list">Sf1a-lectin-his A<sup>r</sup> 8/24</li>
-                 </ul>
-        </ul>
-       <p class="note-title">Protein purification</p>
-           <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-               <li class="list">Sample</li>
-                 <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                      <li class="list">Hv1a-lectin-his</li>
-                 </ul>
-               <li class="list">Collect：<br>pass through solution, wash through solution(W1~3), elution solution(E1~10), filtered NaCl solution, and filtered ddH<sub>2</sub>O solution.</li>
-        </ul>
-       <p class="note-title">Cultivation</p>
-           <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-               <li class="list">Cultivation of sample <i>BL21-Rosetta-gami</i> colonies for protein purification.</li>
-               <li class="list">Sample</li>
-                 <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                      <li class="list">Sf1a-his A<sup>r</sup> 8/17</li>
-                      <li class="list">OAIP-his A<sup>r</sup> 8/17</li>
-                 </ul>
-        </ul>
-       <p class="note-title">IPTG induction</p>
-           <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-               <li class="list">Induce the mass protein expression for SDS-PAGE.</li>
-               <li class="list">The protein would form inclusion body. Therefore, it was devided into two parts, the urea and control.</li>
-               <li class="list">Sample</li>
-                 <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                      <li class="list">Hv1a-his A<sup>r</sup> 8/26</li>
-                      <li class="list">Hv1a-lectin-his A<sup>r</sup> 8/26</li>
-                      <li class="list">Sf1a-lectin-his A<sup>r</sup> 8/26</li>
-                      <li class="list">OAIP-lectin-his A<sup>r</sup> 8/26</li>
-                 </ul>
-               <li class="list">Add 500μM IPTG</li>
-               <li class="list">Take the following samples to go on the experience first. The others storage in 4℃ refrigerator.</li>
-                 <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                      <li class="list">Hv1a-his A<sup>r</sup> 8/26</li>
-                      <li class="list">Hv1a-lectin-his A<sup>r</sup> 8/26</li>
-                 </ul>
-        </ul>
-       <p class="note-title">SDS-PAGE</p>
-           <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-               <li class="list">Check the protein in pellet or in solution.</li>
-               <li class="list">Sample</li>
-               <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                      <li class="list">Hv1a-his A<sup>r</sup> 8/26(with urea)</li>
-                      <li class="list">Hv1a-his A<sup>r</sup> 8/26</li>
-                      <li class="list">Hv1a-lectin-his A<sup>r</sup> 8/26(with urea)</li>
-                      <li class="list">Hv1a-lectin-his A<sup>r</sup> 8/26</li>
-               </ul>
-               <li class="list">Sonication and the supernatant is loading sample1</li>
-                 <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                      <li class="list">10 seconds on, 40 seconds off and repeat six times.</li>
-                 </ul>
-               <li class="list">Add binding buffer (with/without urea) again and the solution is loading sample 2</li>
-               <li class="list">Sonication again and the supernatant is loading sample 3</li>
-                 <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                      <li class="list">10 seconds on, 40 seconds off and repeat six times.</li>
-                 </ul>
-               <li class="list">Add binding buffer (with/without urea) again and the solution is loading sample4</li>
-               <li class="list">Loading sample</li>
-                 <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                      <li class="list">Hv1a-his (with urea) 1~4</li>
-                      <li class="list">Hv1a-his 1~4</li>
-                      <li class="list">Hv1a-lectin-his (with urea) 1~4</li>
-                      <li class="list">Hv1a-lectin-his 1~4</li>
-                 </ul>
-        </ul>
-       <p class="note-title">IPTG induction</p>
-           <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-               <li class="list">Induce the mass protein expression for SDS-PAGE.</li>
-               <li class="list">The protein would form inclusion body. Therefore, it was devided into two parts, the urea and control.</li>
-               <li class="list">Sample</li>
-                 <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                      <li class="list">Hv1a-his A<sup>r</sup> 8/26</li>
-                      <li class="list">Hv1a-lectin-his A<sup>r</sup> 8/26</li>
-                      <li class="list">Sf1a-lectin-his A<sup>r</sup> 8/26</li>
-                      <li class="list">OAIP-lectin-his A<sup>r</sup> 8/26</li>
-                 </ul>
-               <li class="list">Add 500μM IPTG and test OD<sub>600</sub> to approximately 1.050</li>
-        </ul>
-       <p class="note-title">SDS-PAGE</p>
-           <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-               <li class="list">Check the protein in pellet or in solution.</li>
-               <li class="list">Sample</li>
-               <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                      <li class="list">Hv1a-his A<sup>r</sup> 8/26(with urea)</li>
-                      <li class="list">Hv1a-his A<sup>r</sup> 8/26</li>
-                      <li class="list">Hv1a-lectin-his A<sup>r</sup> 8/26(with urea)</li>
-                      <li class="list">Hv1a-lectin-his A<sup>r</sup> 8/26</li>
-                      <li class="list">Sf1a-lectin-his A<sup>r</sup> 8/26(with urea)</li>
-                      <li class="list">Sf1a-lectin-his A<sup>r</sup> 8/26</li>
-                      <li class="list">OAIPlectin-his A<sup>r</sup> 8/26(with urea)</li>
-                      <li class="list">OAIP-lectin-his A<sup>r</sup> 8/26</li>
-               </ul>
-               <li class="list">Certificate the sample, add binding buffer (with/without urea), PMSF</li>
-               <li class="list">Sonication and the supernatant is loading sample1</li>
-                 <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                      <li class="list">10 seconds on, 40 seconds off and repeat six times.</li>
-                 </ul>
-               <li class="list">Add binding buffer (with/without urea) again and the solution is loading sample 2</li>
-               <li class="list">Sonication again and the supernatant is loading sample 3</li>
-                 <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                      <li class="list">10 seconds on, 40 seconds off and repeat six times.</li>
-                 </ul>
-               <li class="list">Add binding buffer (with/without urea) again and the solution is loading sample4</li>
-               <li class="list">Loading sample</li>
-                 <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                      <li class="list">Hv1a-his (with urea) 1~4</li>
-                      <li class="list">Hv1a-his 1~4</li>
-                      <li class="list">Hv1a-lectin-his (with urea) 1~4</li>
-                      <li class="list">Hv1a-lectin-his 1~4</li>
-                      <li class="list">Sf1a-lectin-his (with urea) 1~4</li>
-                      <li class="list">Sf1a-lectin-his 1~4</li>
-                      <li class="list">OAIPlectin-his (with urea) 1~4</li>
-                      <li class="list">OAIP-lectin-his 1~4</li>
-                 </ul>
-        </ul>
       </article>
       <article id="August-27" class="note-item">
          <h2>August 27</h2><hr>
-        <p class="note-title">Cultivation</p>
+        <p class="note-title">None</p>
-           <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-               <li class="list">Cultivation of sample BL21-Rosetta-gami colonies for protein purification.</li>
-               <li class="list">Sample</li>
-                 <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                      <li class="list">Hv1a-his A<sup>r</sup> </li>
-                      <li class="list">Hv1a-lectin-his  A<sup>r</sup> </li>
-                 </ul>
-        </ul>
       </article>
       <article id="August-28" class="note-item">
          <h2>August 28</h2><hr>
-        <p class="note-title">Protein purification</p>
+        <p class="note-title">None</p>
-           <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-               <li class="list">The buffer contains the urea</li>
-               <li class="list">Sample(8/27)</li>
-                 <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                      <li class="list">Hv1a-his</li>
-                      <li class="list">Hv1a-lectin-his</li>
-                 </ul>
-               <li class="list">Collect：<br>pass through solution, wash through solution(W1~3), elution solution(E1~10), filtered NaCl solution, and filtered ddH<sub>2</sub>O solution.</li>
-        </ul>
-       <p class="note-title">SDS-PAGE</p>
-           <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-               <li class="list">Check the purification protein.</li>
-               <li class="list">Sample(8/28)</li>
-                 <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                      <li class="list">Hv1a-his</li>
-                      <li class="list">Hv1a-lectin-his</li>
-                 </ul>
-        </ul>
-       <p class="note-title">Cultivation</p>
-           <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-               <li class="list">Cultivation of sample BL21-Rosetta-gami colonies for protein purification.</li>
-               <li class="list">Sample</li>
-                 <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                      <li class="list">Sf1a-his A<sup>r</sup> 8/17</li>
-                 </ul>
-        </ul>
       </article>
       <article id="August-29" class="note-item">
          <h2>August 29</h2><hr>
-        <p class="note-title">Protein purification</p>
+        <p class="note-title">None</p>
-           <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-               <li class="list">The buffer contains the urea</li>
-               <li class="list">Sample(8/28)</li>
-                 <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                      <li class="list">Sf1a-lectin-his</li>
-                 </ul>
-               <li class="list">Collect：<br>pass through solution, wash through solution(W1~3), elution solution(E1~10), filtered NaCl solution, and filtered ddH<sub>2</sub>O solution.</li>
-        </ul>
-       <p class="note-title">SDS-PAGE</p>
-           <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-               <li class="list">Check the purification protein.</li>
-               <li class="list">Sample(8/29)</li>
-                 <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                      <li class="list">Sf1a-lectin-his</li>
-                 </ul>
-        </ul>
-       <p class="note-title">Cultivation</p>
-           <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-               <li class="list">Cultivation of sample BL21-Rosetta-gami colonies for protein purification.</li>
-               <li class="list">Sample</li>
-                 <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                      <li class="list">Sf1a-his A<sup>r</sup> 8/17</li>
-                 </ul>
-        </ul>
-       <p class="note-title">Cultivation</p>
-           <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-               <li class="list">Cultivation of sample <i>BL21-Rosetta-gami</i> colonies for protein purification.</li>
-               <li class="list">Sample</li>
-                 <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                      <li class="list">Hv1a-his A<sup>r</sup> 8/15</li>
-                 </ul>
-        </ul>
-       <p class="note-title">Cultivation</p>
-           <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-               <li class="list">Cultivation of sample <i>BL21-Rosetta-gami</i> colonies for protein purification.</li>
-               <li class="list">Sample</li>
-                 <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                      <li class="list">Hv1a-lectin-his A<sup>r</sup> 8/15</li>
-                      <li class="list">Sf1a-lectin-his A<sup>r</sup> 8/15</li>
-                      <li class="list">OAIP-lectin-his A<sup>r</sup> 8/15</li>
-                 </ul>
-        </ul>
       </article>
       <article id="August-30" class="note-item">
          <h2>August 30</h2><hr>
-        <p class="note-title">IPTG induction</p>
+        <p class="note-title">None</p>
-           <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-               <li class="list">Induce the mass protein expression for SDS-PAGE.</li>
-               <li class="list">The protein would form inclusion body. Therefore, it was devided into two parts, the urea and control.</li>
-               <li class="list">Sample</li>
-               <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                      <li class="list">Hv1a-lectin-his A<sup>r</sup> 8/29</li>
-                      <li class="list">Sf1a-lectin-his A<sup>r</sup> 8/29</li>
-                      <li class="list">OAIP-lectin-his A<sup>r</sup> 8/29</li>
-               </ul>
-               <li class="list">Refresh to approximately OD<sub>600</sub> 0.6</li>
-               <li class="list">Add 500μM IPTG and cultivate for 4hr</li>
-        </ul>
-       <p class="note-title">Protein purification</p>
-           <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-               <li class="list">The buffer contains the urea</li>
-               <li class="list">Sample(8/30)</li>
-                 <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                      <li class="list">Sf1a-lectin-his</li>
-                      <li class="list">Hv1a-lectin-his</li>
-                 </ul>
-               <li class="list">Collect：<br>pass through solution, wash through solution(W1~3), elution solution(E1~10), filtered NaCl solution, and filtered ddH<sub>2</sub>O solution.</li>
-        </ul>
-       <p class="note-title">SDS-PAGE</p>
-           <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-               <li class="list">Check the purification protein.</li>
-               <li class="list">Sample(8/29)</li>
-                 <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                      <li class="list">Sf1a-lectin-his</li>
-                      <li class="list">Hv1a-lectin-his</li>
-                 </ul>
-        </ul>
-       <p class="note-title">PCR</p>
-           <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-               <li class="list">amplify the insert gene</li>
-               <li class="list">Sample(8/30)</li>
-                 <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                      <li class="list">Hv1a-his</li>
-                      <li class="list">Hv1a-lectin-his</li>
-                      <li class="list">Sf1a-his</li>
-                      <li class="list">Sf1a-lectin-his</li>
-                      <li class="list">OAIP-his</li>
-                      <li class="list">OAIP-lectin-his</li>
-                 </ul>
-        </ul>
-       <p class="note-title">Electrophoresis</p>
-           <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-               <li class="list">Electrophoresis of PCR product</li>
-               <li class="list">Sample(8/30 PCR product)</li>
-                 <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                      <li class="list">Hv1a-his</li>
-                      <li class="list">Hv1a-lectin-his</li>
-                      <li class="list">Sf1a-his</li>
-                      <li class="list">Sf1a-lectin-his</li>
-                      <li class="list">OAIP-his</li>
-                      <li class="list">OAIP-lectin-his</li>
-                 </ul>
-        </ul>
-       <p class="note-title">Digestion</p>
-           <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-               <li class="list">Digest min for transformation</li>
-               <li class="list">Sample</li>
-                 <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                      <li class="list">Hv1a-his digestion EP</li>
-                      <li class="list">Hv1a-lectin-his digestion ES</li>
-                      <li class="list">Sf1a-his digestion EP</li>
-                      <li class="list">Sf1a-lectin-his digestion ES</li>
-                      <li class="list">OAIP-his digestion EP</li>
-                      <li class="list">OAIP-lectin-his digestion ES</li>
-                 </ul>
-        </ul>
-       <p class="note-title">Transformation</p>
-           <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-               <li class="list">Transformation PSB1C3 to DH5α</li>
-               <li class="list">Sample</li>
-                 <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                      <li class="list">Trans PSB1C3 8/30</li>
-                      <li class="list">Trans PSB1C3 8/30</li>
-                 </ul>
-        </ul>
-       <p class="note-title">Urea test</p>
-           <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-               <li class="list">Check whether protein in inclusion body or not.</li>
-               <li class="list">Sample</li>
-                 <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                      <li class="list">Hv1a-his</li>
-                      <li class="list">Hv1a-his + Urea</li>
-                      <li class="list">Sf1a-his</li>
-                      <li class="list">Sf1a-his + Urea</li>
-                      <li class="list">OAIP-his</li>
-                      <li class="list">OAIP-his + Urea</li>
-                 </ul>
-        </ul>
       </article>
       <article id="August-31" class="note-item">
          <h2>August 31</h2><hr>
-        <p class="note-title">Ligation</p>
+        <p class="note-title">None</p>
-           <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-               <li class="list">Ligase insert gene into the pSB1C3</li>
-               <li class="list">Sample</li>
-                 <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                      <li class="list">Hv1a-his</li>
-                      <li class="list">Hv1a-lectin-his</li>
-                      <li class="list">Sf1a-his</li>
-                      <li class="list">Sf1a-lectin-his</li>
-                      <li class="list">OAIP-his</li>
-                      <li class="list">OAIP-lectin-his</li>
-                 </ul>
-        </ul>
-       <p class="note-title">Transformation</p>
-           <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-               <li class="list">Transformation of ligated product to DH5α</li>
-               <li class="list">Sample(8/31 Ligation product)</li>
-                 <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                      <li class="list">Hv1a-his</li>
-                      <li class="list">Hv1a-lectin-his</li>
-                      <li class="list">Sf1a-his</li>
-                      <li class="list">Sf1a-lectin-his</li>
-                      <li class="list">OAIP-his</li>
-                      <li class="list">OAIP-lectin-his</li>
-                 </ul>
-        </ul>
-       <p class="note-title">Dialysis and refolding</p>
-           <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-               <li class="list">Dialysis and refolding for insoluble protein</li>
-               <li class="list">Sample(8/8)</li>
-                 <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                      <li class="list">Hv1a-lectin-his (8/30)</li>
-                      <li class="list">Sf1a-lectin-his(8/30)</li>
-                      <li class="list">OAIP-lectin-his(8/30)</li>
-                      <li class="list">Hv1a-his(8/28)</li>
-                 </ul>
-        </ul>
-       <p class="note-title">SDS-PAGE</p>
-           <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-               <li class="list">Check the purification protein.</li>
-               <li class="list">Sample(8/30)</li>
-                 <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                      <li class="list">Sf1a-lectin-his</li>
-                      <li class="list">Hv1a-lectin-his</li>
-                 </ul>
-        </ul>
       </article>
@@ Line 2,851: / Line 1,414: @@
       <article id="September-1" class="note-item">
          <h2>September 1</h2><hr>
-        <p class="note-title">Protein purification</p>
+        <p class="note-title">None</p>
-           <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-               <li class="list">The buffer contains the urea</li>
-               <li class="list">Sample(8/30)</li>
-                 <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                      <li class="list">OAIP-lectin-his</li>
-                      <li class="list">Hv1a-his</li>
-                 </ul>
-               <li class="list">Collect：<br>pass through solution, wash through solution(W1~3), elution solution(E1~10), filtered NaCl solution, and filtered ddH<sub>2</sub>O solution.</li>
-        </ul>
-       <p class="note-title">SDS-PAGE</p>
-           <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-               <li class="list">Check the purification protein.</li>
-               <li class="list">Sample(8/30)</li>
-                 <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                      <li class="list">OAIP-lectin-his</li>
-                      <li class="list">Hv1a-his</li>
-                 </ul>
-        </ul>
-       <p class="note-title">Cultivation</p>
-           <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-               <li class="list">Cultivation of sample BL21-Rosetta-gami colonies for protein purification.</li>
-               <li class="list">Sample</li>
-                 <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                      <li class="list">Hv1a-his A<sup>r</sup> </li>
-                      <li class="list">Hv1a-lectin-his  A<sup>r</sup> </li>
-                      <li class="list">OAIP-his A<sup>r</sup> </li>
-                      <li class="list">Sf1a-his A<sup>r</sup> </li>
-                 </ul>
-        </ul>
-       <p class="note-title">IPTG Induction</p>
-           <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-               <li class="list">Prepare for the purify the protein from sample <i>BL21-Rosetta-gami</i> colonies.</li>
-               <li class="list">Sample</li>
-                 <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                      <li class="list">Hv1a-his A<sup>r</sup> </li>
-                      <li class="list">OAIP-his A<sup>r</sup> </li>
-                      <li class="list">Sf1a-his A<sup>r</sup> </li>
-                 </ul>
-               <li class="list">Add IPTG 500μM and test OD<sub>600</sub> after 4hr.</li>
-        </ul>
-       <p class="note-title">Sonication</p>
-           <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-               <li class="list">Sample</li>
-                 <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                      <li class="list">Hv1a-his A<sup>r</sup> </li>
-                      <li class="list">OAIP-his A<sup>r</sup> </li>
-                      <li class="list">Sf1a-his A<sup>r</sup> </li>
-                 </ul>
-        </ul>
-       <p class="note-title">Cultivation</p>
-           <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-               <li class="list">Cultivation of sample <i>BL21-Rosetta-gami</i> colonies for protein purification.</li>
-               <li class="list">Sample</li>
-                 <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                      <li class="list">Sf1a-lectin-his A<sup>r</sup></li>
-                      <li class="list">OAIP-lectin-his A<sup>r</sup></li>
-                 </ul>
-        </ul>
-       <p class="note-title">Sonication</p>
-           <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-               <li class="list">Sample</li>
-                 <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                      <li class="list">Sf1a-lectin-his A<sup>r</sup></li>
-                      <li class="list">OAIP-lectin-his A<sup>r</sup></li>
-                 </ul>
-        </ul>
       </article>
       <article id="September-2" class="note-item">
          <h2>September 2</h2><hr>
-       <p class="note-title">Protein purification</p>
+         <p class="note-title">None</p>
-           <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-               <li class="list">The buffer contains the urea</li>
-               <li class="list">Sample</li>
-                 <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                      <li class="list">Sf1a-lectin-his A<sup>r</sup></li>
-                      <li class="list">OAIP-lectin-his A<sup>r</sup></li>
-                 </ul>
-               <li class="list">Collect：<br>pass through solution, wash through solution(W1~3), elution solution(E1~10), filtered NaCl solution, and filtered ddH<sub>2</sub>O solution.</li>
-         </ul>
-       <p class="note-title">Protein quantification Braford method</p>
-           <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-               <li class="list">Sample</li>
-                 <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                      <li class="list">Hv1a-his</li>
-                      <li class="list">Hv1a-lectin-his A<sup>r</sup></li>
-                      <li class="list">Sf1a-lectin-his A<sup>r</sup></li>
-                      <li class="list">OAIP-lectin-his A<sup>r</sup></li>
-                 </ul>
-        </ul>
-       <p class="note-title">Dialysis and refolding</p>
-           <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-               <li class="list">Dialysis and refolding for insoluble protein</li>
-               <li class="list">Sample</li>
-                 <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                      <li class="list">Hv1a-his</li>
-                      <li class="list">Hv1a-lectin-his</li>
-                      <li class="list">Sf1a-his</li>
-                      <li class="list">Sf1a-lectin-his</li>
-                      <li class="list">OAIP-his</li>
-                      <li class="list">OAIP-lectin-his</li>
-                 </ul>
-        </ul>
       </article>
       <article id="September-3" class="note-item">
          <h2>September 3</h2><hr>
-        <p class="note-title">Protein purification</p>
+        <p class="note-title">None</p>
-           <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-               <li class="list">Sample</li>
-                 <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                      <li class="list">Sf1a-lectin-his A<sup>r</sup></li>
-                      <li class="list">OAIP-lectin-his A<sup>r</sup></li>
-                 </ul>
-               <li class="list">Collect：<br>pass through solution, wash through solution(W1~3), elution solution(E1~10), filtered NaCl solution, and filtered ddH<sub>2</sub>O solution.</li>
-        </ul>
-       <p class="note-title">Protein quantification Braford method</p>
-           <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-               <li class="list">The buffer contains the urea</li>
-                 <li class="list">Sample</li>
-                 <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                      <li class="list">Hv1a-lectin-his A<sup>r</sup></li>
-                      <li class="list">Sf1a-lectin-his A<sup>r</sup></li>
-                      <li class="list">OAIP-lectin-his A<sup>r</sup></li>
-                 </ul>
-        </ul>
-       <p class="note-title">Concentrate protein</p>
-           <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-               <li class="list">Increase the concentration of protein</li>
-               <li class="list">Sample</li>
-                 <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                      <li class="list">Hv1a-lectin-his A<sup>r</sup></li>
-                      <li class="list">Sf1a-lectin-his A<sup>r</sup></li>
-                      <li class="list">OAIP-lectin-his A<sup>r</sup></li>
-                 </ul>
-        </ul>
       </article>
       <article id="September-4" class="note-item">
          <h2>September 4</h2><hr>
-         <p class="note-title">Protein purification</p>
+         <p class="note-title">None</p>
-           <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-               <li class="list">The buffer contains the urea</li>
-               <li class="list">Sample</li>
-                 <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                      <li class="list">Sf1a-lectin-his A<sup>r</sup></li>
-                      <li class="list">OAIP-lectin-his A<sup>r</sup></li>
-                 </ul>
-               <li class="list">Collect：<br>pass through solution, wash through solution(W1~3), elution solution(E1~10), filtered NaCl solution, and filtered ddH<sub>2</sub>O solution.</li>
-        </ul>
       </article>
       <article id="September-5" class="note-item">
          <h2>September 5</h2><hr>
-        <p class="note-title">IPTG Induction</p>
+        <p class="note-title">None</p>
-           <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
+     </article>
-               <li class="list">Prepare for the purify the protein from sample <i>BL21-Rosetta-gami</i> colonies.</li>
-               <li class="list">Sample</li>
-                 <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                      <li class="list">Hv1a-his</li>
-                      <li class="list">Hv1a-lectin-his A<sup>r</sup></li>
-                      <li class="list">Sf1a-lectin-his A<sup>r</sup></li>
-                      <li class="list">OAIP-lectin-his A<sup>r</sup></li>
-                 </ul>
-               <li class="list">Add IPTG 500μM and test OD<sub>600</sub> after 4hr.</li>
-        </ul>
-       <p class="note-title">Sonication</p>
+     <article id="September-6" class="note-item">
+        <h2>September 6</h2><hr>
+                     <p class="note-title">Transformation</p>
             <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
+               <li class="list">Transforming backbone pTXB1 containing Lacticin Z gene into <i>E. coli</i> ER2566 to produce our target protein as a bio-stimulator.</li>
                 <li class="list">Sample</li>
                   <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                       <li class="list">Hv1a-his</li>
+                       <li class="list">backbone pTXB1 containing Lacticin Z gene mini</li>
-                      <li class="list">Hv1a-lectin-his A<sup>r</sup></li>
-                      <li class="list">Sf1a-lectin-his A<sup>r</sup></li>
-                      <li class="list">OAIP-lectin-his A<sup>r</sup></li>
                   </ul>
-        </ul>
+           </ul>
+        <p class="note-title">Cultivation</p>
-       <p class="note-title">Protein purification</p>
             <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                <li class="list">The buffer contains the urea</li>
+                <li class="list">Cultivate the <i>E. coli</i> ER2566 at 37°C cultivating <i>Bacillus subtilis</i> ER2566 at 37°C carrying the backbone pTXB1 containing Lacticin Z gene to produce our target protein as a bio-stimulator.</li>
                 <li class="list">Sample</li>
                   <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                       <li class="list">Hv1a-his</li>
+                       <li class="list">Fresh colony of <i>E. coli</i> ER2566 carrying the backbone pTXB1 containing Lacticin Z gene from an overnight plate</li>
-                      <li class="list">Hv1a-lectin-his</li>
+                  </ul>
-                  </ul>
+           </ul>
-               <li class="list">Collect：<br>pass through solution, wash through solution(W1~3), elution solution(E1~10), filtered NaCl solution, and filtered ddH<sub>2</sub>O solution.</li>
+         <p class="note-title">IPTG Induction</p>
-         </ul>
-       <p class="note-title">UV test</p>
             <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                <li class="list">Test degradation rate</li>
+                <li class="list">Inducting <i>E. coli</i> ER2566 carrying the backbone pTXB1 containing Lacticin Z gene after cultivating at 37°C to produce our target protein as a bio-stimulator.</li>
                 <li class="list">Sample</li>
                   <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                       <li class="list">Hv1a-his</li>
+                       <li class="list">Culture of <i>E. coli</i> ER2566 carrying the backbone pTXB1 containing Lacticin Z gene</li>
-                      <li class="list">Hv1a-lectin-his A<sup>r</sup></li>
-                      <li class="list">Sf1a-lectin-his A<sup>r</sup></li>
-                      <li class="list">OAIP-lectin-his A<sup>r</sup></li>
                   </ul>
-        </ul>
+            </ul>
-       <p class="note-title">Concentrate protein</p>
-            <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-               <li class="list">Increase the concentration of protein</li>
-               <li class="list">Sample</li>
-                 <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                      <li class="list">Hv1a-lectin-his A<sup>r</sup></li>
-                      <li class="list">Sf1a-lectin-his A<sup>r</sup></li>
-                      <li class="list">OAIP-lectin-his A<sup>r</sup></li>
-                 </ul>
-        </ul>
-       <p class="note-title">Protein quantification Braford method</p>
-           <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-               <li class="list">Sample</li>
-                 <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                      <li class="list">Hv1a-lectin-his A<sup>r</sup></li>
-                      <li class="list">Sf1a-lectin-his A<sup>r</sup></li>
-                      <li class="list">OAIP-lectin-his A<sup>r</sup></li>
-                 </ul>
-        </ul>
-     </article>
-     <article id="September-6" class="note-item">
-        <h2>September 6</h2><hr>
-       <p class="note-title">Protein purification</p>
-           <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-               <li class="list">The buffer contains the urea</li>
-               <li class="list">Sample</li>
-                 <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                      <li class="list">Hv1a-his A<sup>r</sup></li>
-                      <li class="list">Hv1a-lectin-his A<sup>r</sup></li>
-                 </ul>
-               <li class="list">Collect：<br>pass through solution, wash through solution(W1~3), elution solution(E1~10), filtered NaCl solution, and filtered ddH<sub>2</sub>O solution.</li>
-        </ul>
-       <p class="note-title">Protein quantification Braford method</p>
-           <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-               <li class="list">Sample</li>
-                 <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                      <li class="list">Hv1a-his A<sup>r</sup></li>
-                      <li class="list">Hv1a-lectin-his A<sup>r</sup></li>
-                      <li class="list">Sf1a-his A<sup>r</sup></li>
-                      <li class="list">Sf1a-lectin-his A<sup>r</sup></li>
-                      <li class="list">OAIP-his A<sup>r</sup></li>
-                      <li class="list">OAIP-lectin-his A<sup>r</sup></li>
-                 </ul>
-        </ul>
       </article>
       <article id="September-7" class="note-item">
          <h2>September 7</h2><hr>
-       <p class="note-title">Protein purification</p>
+        <p class="note-title">Sonication</p>
             <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                <li class="list">The buffer contains the urea</li>
+                <li class="list">Breaking <i>E. coli</i> ER2566 carrying the backbone pTXB1 containing Lacticin Z gene after cultivating to harvest the protein by sonication.</li>
                 <li class="list">Sample</li>
                   <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                      <li class="list">Sf1a-lectin-his A<sup>r</sup></li>
+                     <li class="list">Culture of <i>E. coli</i> ER2566 carrying the backbone pTXB1 containing Lacticin Z gene induced by IPTG </li>
-                      <li class="list">OAIP-lectin-his A<sup>r</sup></li>
                   </ul>
-               <li class="list">Collect：<br>pass through solution, wash through solution(W1~3), elution solution(E1~10), filtered NaCl solution, and filtered ddH<sub>2</sub>O solution.</li>
          </ul>
+        <p class="note-title">Protein quantification & SDS-PAGE</p>
-       <p class="note-title">SDS-PAGE</p>
             <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                <li class="list">Check the purification protein.</li>
+                <li class="list">Quantificating the protein expressing the backbone pTXB1 containing Lacticin Z gene by Bradford method and running SDS-PAGE to check the protein expression.</li>
                 <li class="list">Sample</li>
                   <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                      <li class="list">OAIP-lectin-his A<sup>r</sup></li>
+                     <li class="list">Protein expressing the backbone pTXB1 containing Lacticin Z gene induced by IPTG </li>
-                      <li class="list">Sf1a-lectin-his A<sup>r</sup></li>
-                 </ul>
-        </ul>
-       <p class="note-title">UV test</p>
-           <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-               <li class="list">Test degradation rate</li>
-               <li class="list">Sample</li>
-                 <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                      <li class="list">Hv1a-his A<sup>r</sup></li>
-                      <li class="list">Hv1a-lectin-his A<sup>r</sup></li>
-                      <li class="list">Sf1a-his A<sup>r</sup></li>
-                      <li class="list">Sf1a-lectin-his A<sup>r</sup></li>
-                      <li class="list">OAIP-his A<sup>r</sup></li>
-                      <li class="list">OAIP-lectin-his A<sup>r</sup></li>
-                 </ul>
-        </ul>
-       <p class="note-title">Cultivation</p>
-           <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-               <li class="list">Cultivation of sample BL21-Rosetta-gami colonies for protein purification.</li>
-               <li class="list">Sample</li>
-                 <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                      <li class="list">Hv1a-his A<sup>r</sup></li>
-                      <li class="list">Hv1a-lectin-his A<sup>r</sup></li>
-                      <li class="list">Sf1a-his A<sup>r</sup></li>
-                      <li class="list">Sf1a-lectin-his A<sup>r</sup></li>
-                      <li class="list">OAIP-his A<sup>r</sup></li>
-                      <li class="list">OAIP-lectin-his A<sup>r</sup></li>
-                 </ul>
-        </ul>
-       <p class="note-title">Cultivation</p>
-           <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-               <li class="list">Cultivation of sample BL21-Rosetta-gami colonies for protein purification.</li>
-               <li class="list">Sample</li>
-                 <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                      <li class="list">Hv1a-his A<sup>r</sup></li>
-                      <li class="list">Hv1a-lectin-his A<sup>r</sup></li>
-                      <li class="list">Sf1a-his A<sup>r</sup></li>
-                      <li class="list">Sf1a-lectin-his A<sup>r</sup></li>
-                      <li class="list">OAIP-his A<sup>r</sup></li>
-                      <li class="list">OAIP-lectin-his A<sup>r</sup></li>
-                 </ul>
-        </ul>
-       <p class="note-title">Sonication</p>
-           <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-               <li class="list">Sample</li>
-                 <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                      <li class="list">Hv1a-his A<sup>r</sup></li>
-                      <li class="list">Hv1a-lectin-his A<sup>r</sup></li>
-                      <li class="list">Sf1a-his A<sup>r</sup></li>
-                      <li class="list">Sf1a-lectin-his A<sup>r</sup></li>
-                      <li class="list">OAIP-his A<sup>r</sup></li>
-                      <li class="list">OAIP-lectin-his A<sup>r</sup></li>
-                 </ul>
-        </ul>
-       <p class="note-title">Protein quantification Braford method</p>
-           <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-               <li class="list">Sample</li>
-                 <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                      <li class="list">Hv1a-his A<sup>r</sup></li>
-                      <li class="list">Hv1a-lectin-his A<sup>r</sup></li>
-                      <li class="list">Sf1a-his A<sup>r</sup></li>
-                      <li class="list">Sf1a-lectin-his A<sup>r</sup></li>
-                      <li class="list">OAIP-his A<sup>r</sup></li>
-                      <li class="list">OAIP-lectin-his A<sup>r</sup></li>
                   </ul>
          </ul>
@@ Line 3,206: / Line 1,487: @@
       <article id="September-8" class="note-item">
          <h2>September 8</h2><hr>
-        <p class="note-title">Protein purification</p>
+        <p class="note-title">None</p>
-           <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-               <li class="list">The buffer contains the urea</li>
-               <li class="list">Sample</li>
-                 <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                      <li class="list">Hv1a-his A<sup>r</sup></li>
-                      <li class="list">Hv1a-lectin-his A<sup>r</sup></li>
-                 </ul>
-               <li class="list">Collect：<br>pass through solution, wash through solution(W1~3), elution solution(E1~10), filtered NaCl solution, and filtered ddH<sub>2</sub>O solution.</li>
-        </ul>
-       <p class="note-title">SDS-PAGE</p>
-           <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-               <li class="list">Check the purification protein.</li>
-               <li class="list">Sample</li>
-                 <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                      <li class="list">Hv1a-his A<sup>r</sup></li>
-                      <li class="list">Hv1a-lectin-his A<sup>r</sup></li>
-                 </ul>
-        </ul>
-       <p class="note-title">Cultivation and IPTG induction</p>
-           <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-               <li class="list">Cultivation of sample BL21-Rosetta-gami colonies for protein purification.</li>
-               <li class="list">Sample</li>
-                 <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                      <li class="list">Hv1a-his A<sup>r</sup></li>
-                      <li class="list">Hv1a-lectin-his A<sup>r</sup></li>
-                      <li class="list">Sf1a-his A<sup>r</sup></li>
-                      <li class="list">Sf1a-lectin-his A<sup>r</sup></li>
-                      <li class="list">OAIP-his A<sup>r</sup></li>
-                      <li class="list">OAIP-lectin-his A<sup>r</sup></li>
-                 </ul>
-               <li class="list">Refresh to approximately OD<sub>600</sub> 0.7</li>
-               <li class="list">Add 1000μM IPTG and test OD<sub>600</sub> </li>
-                  <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                      <li class="list">Hv1a-his:0.772</li>
-                      <li class="list">Hv1a-lectin-his:0.631</li>
-                      <li class="list">Sf1a-his:0.567</li>
-                      <li class="list">Sf1a-lectin-his:0.78</li>
-                      <li class="list">OAIP-his:0.851</li>
-                      <li class="list">OAIP-lectin-his:0.819</li>
-                 </ul>
-              <li class="list">Certification, and add 2mL binding buffer and 200μM PMSF for sonication. Repeat three times.</li>
-        </ul>
-       <p class="note-title">Sonication</p>
-           <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-               <li class="list">Sample</li>
-                 <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                      <li class="list">Hv1a-his A<sup>r</sup></li>
-                      <li class="list">Hv1a-lectin-his A<sup>r</sup></li>
-                      <li class="list">Sf1a-his A<sup>r</sup></li>
-                      <li class="list">Sf1a-lectin-his A<sup>r</sup></li>
-                      <li class="list">OAIP-his A<sup>r</sup></li>
-                      <li class="list">OAIP-lectin-his A<sup>r</sup></li>
-                 </ul>
-        </ul>
-       <p class="note-title">Sonication</p>
-           <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-               <li class="list">Sample</li>
-                 <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                      <li class="list">Hv1a-his A<sup>r</sup></li>
-                      <li class="list">Hv1a-lectin-his A<sup>r</sup></li>
-                      <li class="list">Sf1a-his A<sup>r</sup></li>
-                      <li class="list">Sf1a-lectin-his A<sup>r</sup></li>
-                      <li class="list">OAIP-his A<sup>r</sup></li>
-                      <li class="list">OAIP-lectin-his A<sup>r</sup></li>
-                 </ul>
-        </ul>
-       <p class="note-title">Protein purification</p>
-           <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-               <li class="list">The buffer contains the urea</li>
-               <li class="list">Sample</li>
-                 <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                      <li class="list">Sf1a-his A<sup>r</sup></li>
-                      <li class="list">Sf1a-lectin-his A<sup>r</sup></li>
-                      <li class="list">OAIP-his A<sup>r</sup></li>
-                      <li class="list">OAIP-lectin-his A<sup>r</sup></li>
-                 </ul>
-               <li class="list">Collect：<br>pass through solution, wash through solution(W1~3), elution solution(E1~10), filtered NaCl solution, and filtered ddH<sub>2</sub>O solution.</li>
-        </ul>
-       <p class="note-title">SDS-PAGE</p>
-           <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-               <li class="list">Check the purification protein.</li>
-               <li class="list">Sample</li>
-                 <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                      <li class="list">Sf1a-his A<sup>r</sup></li>
-                      <li class="list">Sf1a-lectin-his A<sup>r</sup></li>
-                      <li class="list">OAIP-his A<sup>r</sup></li>
-                      <li class="list">OAIP-lectin-his A<sup>r</sup></li>
-                 </ul>
-        </ul>
       </article>
       <article id="September-9" class="note-item">
          <h2>September 9</h2><hr>
-        <p class="note-title">Protein purification</p>
+        <p class="note-title">None</p>
-           <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-               <li class="list">The buffer contains the urea</li>
-               <li class="list">Sample</li>
-                 <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                      <li class="list">Sf1a-his A<sup>r</sup></li>
-                      <li class="list">Sf1a-lectin-his A<sup>r</sup></li>
-                      <li class="list">OAIP-his A<sup>r</sup></li>
-                      <li class="list">OAIP-lectin-his A<sup>r</sup></li>
-                 </ul>
-               <li class="list">Collect：<br>pass through solution, wash through solution(W1~3), elution solution(E1~10), filtered NaCl solution, and filtered ddH<sub>2</sub>O solution.</li>
-        </ul>
-       <p class="note-title">SDS-PAGE</p>
-           <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-               <li class="list">Check the purification protein.</li>
-               <li class="list">Sample</li>
-                 <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                      <li class="list">Sf1a-his A<sup>r</sup></li>
-                      <li class="list">Sf1a-lectin-his A<sup>r</sup></li>
-                      <li class="list">OAIP-his A<sup>r</sup></li>
-                      <li class="list">OAIP-lectin-his A<sup>r</sup></li>
-                 </ul>
-        </ul>
       </article>
@@ Line 3,343: / Line 1,502: @@
          <p class="note-title">None</p>
       </article>
       <article id="September-11" class="note-item">
          <h2>September 11</h2><hr>
-        <p class="note-title">Dialysis protein</p>
+        <p class="note-title">None</p>
-           <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-               <li class="list">Dialysis and refolding for insoluble protein</li>
-               <li class="list">Sample</li>
-                 <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                      <li class="list">Hv1a-his  E1-E3</li>
-                      <li class="list">Hv1a-lectin-his  E1-E5</li>
-                      <li class="list">Sf1a-his  E1-E5</li>
-                      <li class="list">Sf1a-lectin-his E1-E4</li>
-                      <li class="list">OAIP-his  E1-E3</li>
-                      <li class="list">OAIP-lectin-his  E1-E2</li>
-                 </ul>
-        </ul>
       </article>
       <article id="September-12" class="note-item">
          <h2>September 12</h2><hr>
-        <p class="note-title">Protein quantification Braford method</p>
+        <p class="note-title">None</p>
-           <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-               <li class="list">Sample</li>
-                 <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                      <li class="list">Hv1a-his A<sup>r</sup></li>
-                      <li class="list">Hv1a-lectin-his A<sup>r</sup></li>
-                      <li class="list">Sf1a-his A<sup>r</sup></li>
-                      <li class="list">Sf1a-lectin-his A<sup>r</sup></li>
-                      <li class="list">OAIP-his A<sup>r</sup></li>
-                      <li class="list">OAIP-lectin-his A<sup>r</sup></li>
-                 </ul>
-        </ul>
       </article>
@@ Line 3,380: / Line 1,517: @@
          <p class="note-title">None</p>
       </article>
       <article id="September-14" class="note-item">
          <h2>September 14</h2><hr>
          <p class="note-title">None</p>
       </article>
       <article id="September-15" class="note-item">
          <h2>September 15</h2><hr>
          <p class="note-title">None</p>
       </article>
       <article id="September-16" class="note-item">
          <h2>September 16</h2><hr>
          <p class="note-title">None</p>
       </article>
       <article id="September-17" class="note-item">
          <h2>September 17</h2><hr>
-        <p class="note-title">Dialysis protein</p>
+        <p class="note-title">None</p>
-           <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-               <li class="list">Dialysis and refolding for insoluble protein</li>
-               <li class="list">Sample</li>
-                 <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                      <li class="list">Hv1a-his  E2-E3</li>
-                      <li class="list">Hv1a-lectin-his  E1-E6</li>
-                      <li class="list">Sf1a-his  E1-E4</li>
-                      <li class="list">Sf1a-lectin-his E2-E4</li>
-                      <li class="list">OAIP-his  E2-E4</li>
-                      <li class="list">OAIP-lectin-his  E2-E3</li>
-                 </ul>
-        </ul>
       </article>
@@ Line 3,413: / Line 1,542: @@
          <p class="note-title">None</p>
       </article>
       <article id="September-19" class="note-item">
          <h2>September 19</h2><hr>
          <p class="note-title">None</p>
       </article>
       <article id="September-20" class="note-item">
          <h2>September 20</h2><hr>
          <p class="note-title">None</p>
       </article>
       <article id="September-21" class="note-item">
          <h2>September 21</h2><hr>
-         <p class="note-title">None</p>
+         <p class="note-title">Functional analysis test</p>
-     </article>
+         <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-     <article id="September-22" class="note-item">
+               <li class="list">Test the function of Lacticin Z (produced by <i>E. coli</i> ER2566) by elisa reader(triplicate)</li>
-         <h2>September 22</h2><hr>
-       <p class="note-title">Protein quantification Braford method</p>
-           <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
                 <li class="list">Sample</li>
                   <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                      <li class="list">Hv1a-his A<sup>r</sup></li>
+                     <li class="list">Control:<br>
-                      <li class="list">Hv1a-lectin-his A<sup>r</sup></li>
+. <i>Bacillus subtilis</i> + Ampicillin<br>
-                      <li class="list">Sf1a-his A<sup>r</sup></li>
+. <i>Bacillus subtilis</i> + Production of the pTXB1 backbone induced by IPTG<br>
-                      <li class="list">Sf1a-lectin-his A<sup>r</sup></li>
+. <i>Bacillus subtilis</i> + LB broth<br>
-                      <li class="list">OAIP-his A<sup>r</sup></li>
+. LB broth for background</li>
-                      <li class="list">OAIP-lectin-his A<sup>r</sup></li>
+                     <li class="list">Test:<br>
+. <i>Bacillus subtilis</i> + Production of the pTXB1 backbone containing Lacticin Z gene induced by IPTG</li><br>
                   </ul>
          </ul>
+     </article>
+     <article id="September-22" class="note-item">
+        <h2>September 22</h2><hr>
+          <p class="note-title">None</p>
       </article>
       <article id="September-23" class="note-item">
          <h2>September 23</h2><hr>
-       <p class="note-title">Dialysis protein</p>
+          <p class="note-title">None</p>
-           <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-               <li class="list">Remove urea</li>
-               <li class="list">Sample</li>
-                 <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                      <li class="list">Hv1a-his E1-E3</li>
-                      <li class="list">Hv1a-lectin-his E1-E6</li>
-                      <li class="list">Sf1a-his E1-E4</li>
-                      <li class="list">Sf1a-lectin-his E1-E6</li>
-                      <li class="list">OAIP-his E1-E3</li>
-                      <li class="list">OAIP-lectin-his E1-E6</li>
-                 </ul>
-        </ul>
-               <p class="note-title">SDS-PAGE</p>
-           <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-               <li class="list">Check the purification protein again</li>
-               <li class="list">Sample</li>
-                 <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                      <li class="list">Hv1a-his </li>
-                      <li class="list">Hv1a-lectin-his (no β-me)</li>
-                      <li class="list">Sf1a-his </li>
-                      <li class="list">Sf1a-lectin-his (no β-me)</li>
-                      <li class="list">OAIP-his </li>
-                      <li class="list">OAIP-lectin-his (no β-me)</li>
-                 </ul>
-        </ul>
-               <p class="note-title">Electrophoresis</p>
-           <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-               <li class="list">Electrophoresis of composite parts again</li>
-               <li class="list">Sample(8/30 PCR product)</li>
-                 <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                      <li class="list">Hv1a-his</li>
-                      <li class="list">Hv1a-lectin-his</li>
-                      <li class="list">Sf1a-his</li>
-                      <li class="list">Sf1a-lectin-his</li>
-                      <li class="list">OAIP-his</li>
-                      <li class="list">OAIP-lectin-his</li>
-                 </ul>
-        </ul>
       </article>
       <article id="September-24" class="note-item">
          <h2>September 24</h2><hr>
-               <p class="note-title">Electrophoresis</p>
+          <p class="note-title">None</p>
-           <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-               <li class="list">Electrophoresis of composite parts again</li>
-               <li class="list">Sample(8/30 PCR product)</li>
-                 <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                      <li class="list">Hv1a-his</li>
-                      <li class="list">Hv1a-lectin-his</li>
-                      <li class="list">Sf1a-his</li>
-                      <li class="list">Sf1a-lectin-his</li>
-                      <li class="list">OAIP-his</li>
-                      <li class="list">OAIP-lectin-his</li>
-                 </ul>
-        </ul>
       </article>
       <article id="September-25" class="note-item">
          <h2>September 25</h2><hr>
-       <p class="note-title">Protein quantification Braford method</p>
+          <p class="note-title">None</p>
-           <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-               <li class="list">Sample</li>
-                 <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                      <li class="list">Hv1a-his A<sup>r</sup></li>
-                      <li class="list">Hv1a-lectin-his A<sup>r</sup></li>
-                      <li class="list">Sf1a-his A<sup>r</sup></li>
-                      <li class="list">Sf1a-lectin-his A<sup>r</sup></li>
-                      <li class="list">OAIP-his A<sup>r</sup></li>
-                      <li class="list">OAIP-lectin-his A<sup>r</sup></li>
-                 </ul>
-        </ul>
       </article>
@@ Line 3,525: / Line 1,595: @@
          <p class="note-title">None</p>
       </article>
       <article id="September-27" class="note-item">
          <h2>September 27</h2><hr>
          <p class="note-title">None</p>
       </article>
       <article id="September-28" class="note-item">
          <h2>September 28</h2><hr>
-        <p class="note-title">Protein quantification Braford method</p>
+        <p class="note-title">None</p>
-           <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-               <li class="list">Sample</li>
-                 <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                      <li class="list">Hv1a-his A<sup>r</sup></li>
-                      <li class="list">Hv1a-lectin-his A<sup>r</sup></li>
-                      <li class="list">Sf1a-his A<sup>r</sup></li>
-                      <li class="list">OAIP-his A<sup>r</sup></li>
-                 </ul>
-        </ul>
       </article>
       <article id="September-29" class="note-item">
          <h2>September 29</h2><hr>
          <p class="note-title">None</p>
       </article>
       <article id="September-30" class="note-item">
          <h2>September 30</h2><hr>
@@ Line 3,554: / Line 1,619: @@
       <article id="October-1" class="note-item">
-         <h2>October-1</h2><hr>
+         <h2>October 1</h2><hr>
          <p class="note-title">None</p>
       </article>
       <article id="October-2" class="note-item">
          <h2>October 2</h2><hr>
          <p class="note-title">None</p>
       </article>
       <article id="October-3" class="note-item">
          <h2>October 3</h2><hr>
          <p class="note-title">None</p>
       </article>
       <article id="October-4" class="note-item">
          <h2>October 4</h2><hr>
          <p class="note-title">None</p>
       </article>
       <article id="October-5" class="note-item">
          <h2>October 5</h2><hr>
-        <p class="note-title">Cultivation</p>
+        <p class="note-title">Transformation</p>
             <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                <li class="list">For larva test</li>
+                <li class="list">Transforming backbone pTXB1 into <i>E. coli</i> Rosetta-Gami to produce protein as our negative control in verifying the function of our target peptide.</li>
+               <li class="list">Transforming backbone pTXB1 containing bacteriocin gene into <i>E. coli</i> Rosetta-Gami to produce our target protein as a bio-stimulator.</li>
                 <li class="list">Sample</li>
                   <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                       <li class="list">Hv1a-GS linker-lectin  A<sup>r</sup> </li>
+                       <li class="list">backbone pTXB1 mini</li>
+                      <li class="list">backbone pTXB1 containing Enterocin B gene mini</li>
+                      <li class="list">backbone pTXB1 containing Enterocin 96 gene mini</li>
+                      <li class="list">backbone pTXB1 containing Bovicin HJ50 gene mini</li>
+                      <li class="list">backbone pTXB1 containing Durancin TW-49M gene mini</li>
+                      <li class="list">backbone pTXB1 containing Lacticin Z gene mini</li>
+                      <li class="list">backbone pTXB1 containing Leucocyclicin Q gene mini</li>
                   </ul>
          </ul>
@@ Line 3,583: / Line 1,659: @@
       <article id="October-6" class="note-item">
          <h2>October 6</h2><hr>
-         <p class="note-title">None</p>
+         <p class="note-title">Cultivation</p>
+           <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
+               <li class="list">Cultivate the <i>E. coli</i> ER2566 at 37°C cultivating <i>E. coli</i> Rosetta-Gami at 37°C carrying the backbone pTXB1 to produce protein as our negative control in verifying the function of our target peptide.</li>
+               <li class="list">Cultivate the <i>E. coli</i> ER2566 at 37°C cultivating <i>E. coli</i> Rosetta-Gami at 37°C carrying the backbone pTXB1 containing bacteriocin gene produce our target protein as a bio-stimulator.</li>
+               <li class="list">Sample</li>
+                 <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
+                      <li class="list">Fresh colony of <i>E. coli</i> Rosetta-Gami carrying the backbone pTXB1 from an overnight plate</li>
+                      <li class="list">Fresh colony of <i>E. coli</i> Rosetta-Gami carrying the backbone pTXB1 containing Enterocin B gene from an overnight plate</li>
+                      <li class="list">Fresh colony of <i>E. coli</i> Rosetta-Gami carrying the backbone pTXB1 containing Enterocin 96 gene from an overnight plate</li>
+                      <li class="list">Fresh colony of <i>E. coli</i> Rosetta-Gami carrying the backbone pTXB1 containing Leucocyclicin Q gene from an overnight plate</li>
+                 </ul>
+        </ul>
+        <p class="note-title">IPTG Induction</p>
+           <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
+               <li class="list">Cultivate the <i>E. coli</i> ER2566 at 37°C cultivating <i>E. coli</i> Rosetta-Gami at 37°C carrying the backbone pTXB1 to produce protein as our negative control in verifying the function of our target peptide.</li>
+               <li class="list">Inducting <i>E. coli</i> Rosetta-Gami carrying the backbone pTXB1 containing bacteriocin gene after cultivating at 37°C produce our target protein as a bio-stimulator.</li>
+               <li class="list">Sample</li>
+                 <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
+                      <li class="list">Fresh colony of <i>E. coli</i> Rosetta-Gami carrying the backbone pTXB1 from an overnight plate</li>
+                      <li class="list">Culture of <i>E. coli</i> Rosetta-Gami carrying the backbone pTXB1 containing Enterocin B gene</li>
+                      <li class="list">Culture of <i>E. coli</i> Rosetta-Gami carrying the backbone pTXB1 containing Enterocin 96 gene</li>
+                      <li class="list">Culture of <i>E. coli</i> Rosetta-Gami carrying the backbone pTXB1 containing Leucocyclicin Q gene</li>
+                 </ul>
+        </ul>
       </article>
       <article id="October-7" class="note-item">
          <h2>October 7</h2><hr>
-         <p class="note-title">None</p>
+         <p class="note-title">Cultivation</p>
-     </article>
-     <article id="October-8" class="note-item">
-        <h2>October 8</h2><hr>
-       <p class="note-title">Protein quantification Braford method</p>
             <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
+              <li class="list">Cultivate the <i>E. coli</i> ER2566 at 37°C cultivating <i>E. coli</i> Rosetta-Gami at 37°C carrying the backbone pTXB1 containing bacteriocin gene produce our target protein as a bio-stimulator.</li>
                 <li class="list">Sample</li>
                   <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                       <li class="list">HLG-1</li>
+                       <li class="list">Fresh colony of <i>E. coli</i> Rosetta-Gami carrying the backbone pTXB1 containing Bovicin HJ50 gene from an overnight plate</li>
-                      <li class="list">HLG-2</li>
+                       <li class="list">Fresh colony of <i>E. coli</i> Rosetta-Gami carrying the backbone pTXB1 containing Durancin TW-49M gene from an overnight plate</li>
-                      <li class="list">Hv1a-his </li>
+                       <li class="list">Fresh colony of <i>E. coli</i> Rosetta-Gami carrying the backbone pTXB1 containing Lacticin Z gene from an overnight plate</li>
-                       <li class="list">Hv1a-lectin-his</li>
-                      <li class="list">Sf1a-his </li>
-                      <li class="list">Sf1a-lectin-his</li>
-                       <li class="list">OAIP-his </li>
-                      <li class="list">OAIP-lectin-his </li>
-                      <li class="list">BL</li>
-                      <li class="list">RG</li>
                   </ul>
          </ul>
+        <p class="note-title">IPTG Induction</p>
-         <p class="note-title">Protein quantification Braford method</p>
             <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
+               <li class="list">Inducting <i>E. coli</i> Rosetta-Gami carrying the backbone pTXB1 containing bacteriocin gene after cultivating at 37°C produce our target protein as a bio-stimulator.</li>
                 <li class="list">Sample</li>
                   <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                       <li class="list">HLG-1</li>
+                       <li class="list">Culture of <i>E. coli</i> Rosetta-Gami carrying the backbone pTXB1 containing Bovicin HJ50 gene</li>
-                      <li class="list">Hv1a-his </li>
+                       <li class="list">Culture of <i>E. coli</i> Rosetta-Gami carrying the backbone pTXB1 containing Durancin TW-49M gene</li>
-                       <li class="list">Hv1a-lectin-his</li>
+                       <li class="list">Culture of <i>E. coli</i> Rosetta-Gami carrying the backbone pTXB1 containing Lacticin Z gene</li>
-                      <li class="list">Sf1a-his </li>
-                      <li class="list">Sf1a-lectin-his</li>
-                       <li class="list">OAIP-his </li>
-                      <li class="list">OAIP-lectin-his </li>
                   </ul>
          </ul>
-     </article>
+         <p class="note-title">Sonication</p>
-     <article id="October-9" class="note-item">
-         <h2>October 9</h2><hr>
-        <p class="note-title">None</p>
-     </article>
-     <article id="October-10" class="note-item">
-        <h2>October 10</h2><hr>
-        <p class="note-title">None</p>
-     </article>
-     <article id="October-11" class="note-item">
-        <h2>October 11</h2><hr>
-       <p class="note-title">Protein quantification Braford method</p>
             <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
+               <li class="list">Breaking <i>E. coli</i> Rosetta-Gami carrying the backbone pTXB1 and pTXB1 containing bacteriocin gene after cultivating to harvest the protein by sonication.</li>
                 <li class="list">Sample</li>
                   <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                       <li class="list">HLG-1</li>
+                       <li class="list">Culture of <i>E. coli</i> Rosetta-Gami carrying the backbone pTXB1 induced by IPTG </li>
-                      <li class="list">Hv1a-his </li>
+                       <li class="list">Culture of <i>E. coli</i> Rosetta-Gami carrying the backbone pTXB1 containing Enterocin B gene induced by IPTG</li>
-                       <li class="list">Hv1a-lectin-his</li>
+                       <li class="list">Culture of <i>E. coli</i> Rosetta-Gami carrying the backbone pTXB1 containing Enterocin 96 gene induced by IPTG</li>
-                      <li class="list">Sf1a-his </li>
+                       <li class="list">Culture of <i>E. coli</i> Rosetta-Gami carrying the backbone pTXB1 containing Leucocyclicin Q gene induced by IPTG</li>
-                       <li class="list">Sf1a-lectin-his</li>
-                      <li class="list">OAIP-his </li>
-                       <li class="list">OAIP-lectin-his </li>
-                      <li class="list">RG</li>
                   </ul>
          </ul>
+        <p class="note-title">Protein quantification & SDS-PAGE</p>
-         <p class="note-title">SDS-PAGE</p>
             <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                <li class="list">Check the purification protein.</li>
+                <li class="list">Quantificating the protein expressing the backbone pTXB1 and pTXB1 containing bacteriocin gene by Bradford method and running SDS-PAGE to check the protein expression.</li>
                 <li class="list">Sample</li>
                   <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                       <li class="list">Hv1a(sonication)</li>
+                       <li class="list">Protein expressing the backbone pTXB1 induced by IPTG</li>
-                       <li class="list">Hv1a(purification)</li>
+                       <li class="list">Protein expressing the backbone pTXB1 containing Enterocin B gene induced by IPTG </li>
-                       <li class="list">Sf1a(sonication)</li>
+                       <li class="list">Protein expressing the backbone pTXB1 containing Enterocin 96 gene induced by IPTG </li>
-                       <li class="list">Sf1a(purification)</li>
+                       <li class="list">Protein expressing the backbone pTXB1 containing Leucocyclicin Q gene induced by IPTG </li>
-                      <li class="list">OAIP(sonication)</li>
-                      <li class="list">OAIP(purification)</li>
                   </ul>
          </ul>
+     </article>
-        <p class="note-title">Protein quantification Braford method</p>
+     <article id="October-8" class="note-item">
+        <h2>October 8</h2><hr>
+        <p class="note-title">Sonication</p>
             <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
+               <li class="list">Breaking <i>E. coli</i> Rosetta-Gami carrying the backbone pTXB1 containing bacteriocin gene after cultivating to harvest the protein by sonication.</li>
                 <li class="list">Sample</li>
                   <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                       <li class="list">HLG</li>
+                       <li class="list">Culture of <i>E. coli</i> Rosetta-Gami carrying the backbone pTXB1 containing Bovicin HJ50 gene induced by IPTG</li>
-                      <li class="list">Hv1a-his </li>
+                       <li class="list">Culture of <i>E. coli</i> Rosetta-Gami carrying the backbone pTXB1 containing Durancin TW-49M gene induced by IPTG</li>
-                       <li class="list">Hv1a-lectin-his</li>
+                       <li class="list">Culture of <i>E. coli</i> Rosetta-Gami carrying the backbone pTXB1 containing Lacticin Z gene induced by IPTG</li>
-                       <li class="list">Sf1a-his </li>
+                       <li class="list">Culture of <i>E. coli</i> Rosetta-Gami carrying the backbone pTXB1 containing Leucocyclicin Q gene induced by IPTG</li>
-                      <li class="list">Sf1a-lectin-his</li>
-                       <li class="list">OAIP-his </li>
-                      <li class="list">OAIP-lectin-his </li>
                   </ul>
          </ul>
+        <p class="note-title">Protein quantification & SDS-PAGE</p>
-          <p class="note-title">Protein quantification Braford method</p>
             <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
+               <li class="list">Quantificating the protein expressing the backbone pTXB1 containing bacteriocin gene by Bradford method and running SDS-PAGE to check the protein expression.</li>
                 <li class="list">Sample</li>
                   <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                       <li class="list">HLG(purification)</li>
+                       <li class="list">Protein expressing the backbone pTXB1 containing Bovicin HJ50 gene induced by IPTG </li>
-                       <li class="list">Hv1a-his(purification) </li>
+                       <li class="list">Protein expressing the backbone pTXB1 containing Durancin TW-49M gene induced by IPTG </li>
-                       <li class="list">Hv1a-lectin-his(purification)</li>
+                       <li class="list">Protein expressing the backbone pTXB1 containing Lacticin Z gene induced by IPTG </li>
-                       <li class="list">Sf1a-his(purification)</li>
+                       <li class="list">Protein expressing the backbone pTXB1 containing Leucocyclicin Q gene induced by IPTG </li>
-                      <li class="list">Sf1a-lectin-his(purification)</li>
+                 </ul>
-                      <li class="list">OAIP-his(purification) </li>
-                      <li class="list">OAIP-lectin-his(purification) </li>
-                      <li class="list">Hv1a-his(sonication)</li>
-                      <li class="list">OAIP-his(sonication) </li>
-                      <li class="list">OAIP-lectin-his(sonication) </li>
-                      <li class="list">HGL-his(sonication) </li>
-                      <li class="list">Hv1a-his</li>
-                      <li class="list">OAIP-lectin-his</li>
-                      </ul>
          </ul>
+     </article>
+     <article id="October-9" class="note-item">
+        <h2>October 9</h2><hr>
+        <p class="note-title">Functional analysis test</p>
+        <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
+               <li class="list">Test the function of bacteriocins (produced by <i>E. coli</i> Rosetta-gami) by elisa reader(triplicate)</li>
+               <li class="list">Sample</li>
+                 <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
+                      <li class="list">Control:<br>
+. <i>Bacillus subtilis</i> + Ampicillin<br>
+. <i>Bacillus subtilis</i> + Production of the pTXB1 backbone induced by IPTG<br>
+. <i>Bacillus subtilis</i> + LB broth<br>
+. LB broth for background</li>
+                      <li class="list">Test:<br>
+. <i>Bacillus subtilis</i> + Production of the pTXB1 backbone containing Enterocin B gene induced by IPTG<br>
+. <i>Bacillus subtilis</i> + Production of the pTXB1 backbone containing Enterocin 96 gene induced by IPTG<br>
+. <i>Bacillus subtilis</i> + Production of the pTXB1 backbone containing Lacticin Z gene induced by IPTG<br>
+. <i>Bacillus subtilis</i> + Production of the pTXB1 backbone containing Durancin TW-49M gene induced by IPTG</li>
+                 </ul>
+        </ul>
+     </article>
+     <article id="October-10" class="note-item">
+        <h2>October 10</h2><hr>
+        <p class="note-title">Functional analysis test</p>
+        <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
+               <li class="list">Test the function of bacteriocins (produced by <i>E. coli</i> Rosetta-gami) by elisa reader(triplicate)</li>
+               <li class="list">Sample</li>
+                 <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
+                      <li class="list">Control:<br>
+. <i>Bacillus subtilis</i> + Ampicillin<br>
+. <i>Bacillus subtilis</i> + Production of the pTXB1 backbone induced by IPTG<br>
+. <i>Bacillus subtilis</i> + LB broth<br>
+. LB broth for background</li>
+                      <li class="list">Test:<br>
+. <i>Bacillus subtilis</i> + Production of the pTXB1 backbone containing Bovicin HJ50 gene induced by IPTG<br>
+. <i>Bacillus subtilis</i> + Production of the pTXB1 backbone containing Leucocyclicin Q gene induced by IPTG</li>
+                 </ul>
+        </ul>
+     </article>
+     <article id="October-11" class="note-item">
+        <h2>October 11</h2><hr>
+       <p class="note-title">None</p>
       </article>
       <article id="October-12" class="note-item">
          <h2>October 12</h2><hr>
-         <p class="note-title">None</p>
+         <p class="note-title">Functional analysis test</p>
+        <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
+               <li class="list">Test the function of bacteriocins (produced by <i>E. coli</i> Rosetta-gami) by elisa reader(Dose response assessment)</li>
+               <li class="list">Sample</li>
+                 <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
+                      <li class="list">Control:<br>
+. <i>Bacillus subtilis</i> + Ampicillin<br>
+. <i>Bacillus subtilis</i> + Production of the pTXB1 backbone induced by IPTG<br>
+. <i>Bacillus subtilis</i> + LB broth<br>
+. LB broth for background</li>
+                      <li class="list">Test:<br>
+. <i>Bacillus subtilis</i> + Production of the pTXB1 backbone containing Enterocin B gene induced by IPTG<br>
+. <i>Bacillus subtilis</i> + Production of the pTXB1 backbone containing Enterocin 96 gene induced by IPTG<br>
+. <i>Bacillus subtilis</i> + Production of the pTXB1 backbone containing Lacticin Z gene induced by IPTG<br>
+. <i>Bacillus subtilis</i> + Production of the pTXB1 backbone containing Durancin TW-49M gene induced by IPTG</li>
+                 </ul>
+        </ul>
       </article>
       <article id="October-13" class="note-item">
          <h2>October 13</h2><hr>
          <p class="note-title">None</p>
       </article>
       <article id="October-14" class="note-item">
          <h2>October 14</h2><hr>
          <p class="note-title">None</p>
       </article>
       <article id="October-15" class="note-item">
          <h2>October 15</h2><hr>
-         <p class="note-title">SDS-PAGE</p>
+         <p class="note-title">Functional analysis test</p>
-           <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
+        <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                <li class="list">Check the purification protein.</li>
+                <li class="list">Test the function of bacteriocins (produced by <i>E. coli</i> Rosetta-gami) by elisa reader(Dose response assessment)</li>
                 <li class="list">Sample</li>
                   <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
-                       <li class="list">Hv1a-his</li>
+                       <li class="list">Control:<br>
-                       <li class="list">Hv1a-lectin-his</li>
+. <i>Bacillus subtilis</i> + Ampicillin<br>
-                      <li class="list">Sf1a-his</li>
+. <i>Bacillus subtilis</i> + Production of the pTXB1 backbone induced by IPTG<br>
-                      <li class="list">Sf1a-lectin-his</li>
+. <i>Bacillus subtilis</i> + LB broth<br>
-                       <li class="list">OAIP-his</li>
+. LB broth for background</li>
-                       <li class="list">OAIP-lectin-his</li>
+                       <li class="list">Test:<br>
+. <i>Bacillus subtilis</i> + Production of the pTXB1 backbone containing Bovicin HJ50 gene induced by IPTG<br>
+. <i>Bacillus subtilis</i> + Production of the pTXB1 backbone containing Leucocyclicin Q gene induced by IPTG</li>
+                 </ul>
+        </ul>
+        <p class="note-title">Functional analysis test(inhibition zone)</p>
+        <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
+               <li class="list">Test the function of Enterocin B (produced by <i>E. coli</i> Rosetta-gami) by LB Agar + Ampicillin plate</li>
+               <li class="list">Sample</li>
+                 <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
+                       <li class="list">The <i>Bacillus subtilis</i> on the plate</li>
+                       <li class="list">Control:<br>
+. Ampicillin<br>
+. Production of the pTXB1 backbone induced by IPTG<br>
+                      <li class="list">Test:<br>
+. Production of the pTXB1 backbone containing Enterocin B gene induced by IPTG<br>
                   </ul>
          </ul>
@@ Line 3,729: / Line 1,866: @@
       <article id="October-16" class="note-item">
          <h2>October 16</h2><hr>
-         <p class="note-title">None</p>
+         <p class="note-title">Functional analysis test(inhibition zone)</p>
+        <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
+               <li class="list">Test the function of bacteriocins (produced by <i>E. coli</i> Rosetta-gami) by LB Agar + Ampicillin plate</li>
+               <li class="list">Sample</li>
+                 <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
+                      <li class="list">The <i>Bacillus subtilis</i> on the plate</li>
+                      <li class="list">Control:<br>
+. Ampicillin<br>
+. Production of the pTXB1 backbone induced by IPTG<br>
+                      <li class="list">Test:<br>
+. Production of the pTXB1 backbone containing Bovicin HJ50 gene induced by IPTG<br>
+. Production of the pTXB1 backbone containing Lacticin Z gene induced by IPTG<br>
+                 </ul>
+        </ul>
       </article>
       <article id="October-17" class="note-item">
          <h2>October 17</h2><hr>
-         <p class="note-title">None</p>
+         <p class="note-title">Functional analysis test</p>
+        <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
+               <li class="list">Test the function of bacteriocins (produced by <i>E. coli</i> Rosetta-gami) by elisa reader(Dose response assessment)</li>
+               <li class="list">Sample</li>
+                 <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
+                      <li class="list">Control:<br>
+. <i>Bacillus subtilis</i> + Ampicillin<br>
+. <i>Bacillus subtilis</i> + Production of the pTXB1 backbone induced by IPTG<br>
+. <i>Bacillus subtilis</i> + LB broth<br>
+                      <li class="list">Test:<br>
+. <i>Bacillus subtilis</i> + Production of the pTXB1 backbone containing Durancin TW-49M gene induced by IPTG<br>
+                 </ul>
+        </ul>
       </article>
       <article id="October-18" class="note-item">
          <h2>October 18</h2><hr>
          <p class="note-title">None</p>
       </article>
       <article id="October-19" class="note-item">
          <h2>October 19</h2><hr>

Sun	Mon	Tue	Wed	Thu	Fri	Sat
1	2	3	4	5	6	7
8	9	10	11	12	13	14
15	16	17	18	19	20	21
22	23	24	25	26	27	28
29	30	31

Sun	Mon	Tue	Wed	Thu	Fri	Sat
			1	2	3	4
5	6	7	8	9	10	11
12	13	14	15	16	17	18
19	20	21	22	23	24	25
26	27	28	29	30	31

Sun	Mon	Tue	Wed	Thu	Fri	Sat
						1
2	3	4	5	6	7	8
9	10	11	12	13	14	15
16	17	18	19	20	21	22
23	24	25	26	27	28	29
30

Sun	Mon	Tue	Wed	Thu	Fri	Sat
	1	2	3	4	5	6
7	8	9	10	11	12	13
14	15	16	17	18	19	20
21	22	23	24	25	26	27
28	29	30	31

Difference between revisions of "Team:NCTU Formosa/Notebook"

Latest revision as of 19:43, 17 October 2018

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