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Difference between revisions of "Team:NCTU Formosa/Notebook"
| Line 1,329: | Line 1,329: | ||
<article id="August-24" class="note-item"> | <article id="August-24" class="note-item"> | ||
<h2>August 24</h2><hr> | <h2>August 24</h2><hr> | ||
| − | + | <p class="note-title">Transformation</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">Transforming backbone pTXB1 containing bacteriocin gene into <i>Bacillus subtilis</i> ER2566 to produce our target protein as a biostimulator.</li> | <li class="list">Transforming backbone pTXB1 containing bacteriocin gene into <i>Bacillus subtilis</i> ER2566 to produce our target protein as a biostimulator.</li> | ||
| Line 1,337: | Line 1,337: | ||
<li class="list">backbone pTXB1 containing Durancin gene mini</li> | <li class="list">backbone pTXB1 containing Durancin gene mini</li> | ||
</ul> | </ul> | ||
| + | </ul> | ||
<p class="note-title">Cultivation</p> | <p class="note-title">Cultivation</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;"> | ||
| Line 1,344: | Line 1,345: | ||
<li class="list">Fresh colony of <i>Bacillus subtilis</i> ER2566 carrying the backbone pTXB1 containing Bovicin HJ50 gene from an overnight plate</li> | <li class="list">Fresh colony of <i>Bacillus subtilis</i> ER2566 carrying the backbone pTXB1 containing Bovicin HJ50 gene from an overnight plate</li> | ||
<li class="list">Fresh colony of <i>Bacillus subtilis</i> ER2566 carrying the backbone pTXB1 containing Durancin gene from an overnight plate</li> | <li class="list">Fresh colony of <i>Bacillus subtilis</i> ER2566 carrying the backbone pTXB1 containing Durancin gene from an overnight plate</li> | ||
| − | </ul> | + | </ul> |
| + | </ul> | ||
<p class="note-title">IPTG Induction</p> | <p class="note-title">IPTG Induction</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;"> | ||
| Line 1,353: | Line 1,355: | ||
<li class="list">Culture of <i>Bacillus subtilis</i> carrying the backbone pTXB1 containing Durancin gene</li> | <li class="list">Culture of <i>Bacillus subtilis</i> carrying the backbone pTXB1 containing Durancin gene</li> | ||
</ul> | </ul> | ||
| + | </ul> | ||
</article> | </article> | ||
| Line 1,442: | Line 1,445: | ||
<li class="list">backbone pTXB1 containing Lacticin Z gene mini</li> | <li class="list">backbone pTXB1 containing Lacticin Z gene mini</li> | ||
</ul> | </ul> | ||
| + | </ul> | ||
<p class="note-title">Cultivation</p> | <p class="note-title">Cultivation</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;"> | ||
| Line 1,448: | Line 1,452: | ||
<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">Fresh colony of <i>Bacillus subtilis</i> ER2566 carrying the backbone pTXB1 containing Lacticin Z gene from an overnight plate</li> | <li class="list">Fresh colony of <i>Bacillus subtilis</i> ER2566 carrying the backbone pTXB1 containing Lacticin Z gene from an overnight plate</li> | ||
| − | </ul> | + | </ul> |
| + | </ul> | ||
<p class="note-title">IPTG Induction</p> | <p class="note-title">IPTG Induction</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;"> | ||
| Line 1,456: | Line 1,461: | ||
<li class="list">Culture of <i>Bacillus subtilis</i> carrying the backbone pTXB1 containing Lacticin Z gene</li> | <li class="list">Culture of <i>Bacillus subtilis</i> carrying the backbone pTXB1 containing Lacticin Z gene</li> | ||
</ul> | </ul> | ||
| + | </ul> | ||
</article> | </article> | ||
Revision as of 20:30, 16 October 2018
Wet Lab
July 1
Cloning
- Received and resuspended of the IDT DNA fragments
- Sample
- GS linker+αS1-casein
- Enterocin B
- Enterocin 96
- Bovicin HJ50
- Durancin TW49M
- Lacticin Z
- Leucocyclicin Q
- Received and resuspended of the IDT DNA fragments
July 2
None
July 3
Growth curve exp
- Observe the growth population of Bacillus subtilis in different temperature Condition: 37°C, 32°C, 27°C
July 4
Cloning gene of Curcumin biosensor
- 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.
- Sample
- pet30a backbone
- DNA segment of αS1-casein+GS linker and T7 promoter
July 5
Cloning gene of Curcumin biosensor Expression
- Transforming the ligation product of the Curcumin biosensor 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 biosensor 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
Biosensor sensitivity pretest
- Using DPV(Differential Pulse Voltammetry) to check whether our biosensor can detect curcumin.
1. Add the diluted curcumin samples on our biosensor 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 electrochemical cell. (Fig. 3, left)
5. Use the prototype of electrochemical machine to measure the DPV method
July 12
Biosensor sensitivity assay –Determine Standard Curve and Create the Formula
- Detecting the diluted standard curcumin samples by curcumin biosensor and made the standard curve and doing the polynomial curve fitting.
July 13
Biosensor 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 TW49M + 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 TW49M + pTXB1
August 14
Cloning
- Cultivation
- Miniprep (Purify Plasmid)
- Sample
- Enterocin B + pTXB1
- Enterocin 96 + pTXB1
- Durancin TW49M + 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 TW49M + 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 Bacillus subtilis ER2566 to produce protein as our negative control in verifying the function of our target peptide.
- Transforming backbone pTXB1 containing bacteriocin gene into Bacillus subtilis ER2566 to produce our target protein as a biostimulator.
- 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
- Cultivate the E. coli ER2566 at 37°Ccultivating 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°Ccultivating Bacillus subtilis ER2566 at 37°C carrying the backbone pTXB1 containing bacteriocin gene to produce our target protein as a biostimulator.
- Sample
- Fresh colony of Bacillus subtilis ER2566 carrying the backbone pTXB1 from an overnight plate
- Fresh colony of Bacillus subtilis ER2566 carrying the backbone pTXB1 containing Enterocin B gene from an overnight plate
- Fresh colony of Bacillus subtilis ER2566 carrying the backbone pTXB1 containing Enterocin 96 gene from an overnight plate
- Fresh colony of Bacillus subtilis ER2566 carrying the backbone pTXB1 containing Leucocyclicin Q gene from an overnight plate
- Inducting Bacillus subtilis 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 Bacillus subtilis ER2566 carrying the backbone pTXB1 containing bacteriocin gene after cultivating at 37°C to produce our target protein as a biostimulator.
- Sample
- Culture of Bacillus subtilis carrying the backbone pTXB1
- Culture of Bacillus subtilis carrying the backbone pTXB1 containing Enterocin B gene
- Culture of Bacillus subtilis carrying the backbone pTXB1 containing Enterocin 96 gene
- Culture of Bacillus subtilis carrying the backbone pTXB1 containing Leucocyclicin Q gene
Cultivation
IPTG Induction
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 TW49M + pET30a
- Lacticin Z + pET30a
- Leucocyclicin Q + pET30a
Sonication
- Breaking Bacillus subtilis ER2566 carrying the backbone pTXB1 and pTXB1 containing bacteriocin gene after cultivating to harvest the protein by sonication.
- Sample
- Culture of Bacillus subtilis carrying the backbone pTXB1 induced by IPTG
- Culture of Bacillus subtilis carrying the backbone pTXB1 containing Enterocin B gene induced by IPTG
- Culture of Bacillus subtilis carrying the backbone pTXB1 containing Enterocin 96 gene induced by IPTG
- Culture of Bacillus subtilis 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 TW49M + 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 TW49M + pET30a
- Lacticin Z + pET30a
- Leucocyclicin Q + pET30a
August 22
Cloning
- Sequencing plasmid
- Sample
- Enterocin B + pET30a
- Enterocin 96 + pET30a
- Bovicin HJ50 + pET30a
- Durancin TW49M + pET30a
- Lacticin Z + pET30a
- Leucocyclicin Q + pET30a
August 23
None
August 24
Transformation
- Transforming backbone pTXB1 containing bacteriocin gene into Bacillus subtilis ER2566 to produce our target protein as a biostimulator.
- Sample
- backbone pTXB1 containing Bovicin HJ50 gene mini
- backbone pTXB1 containing Durancin gene mini
Cultivation
- Cultivate the E. coli ER2566 at 37°Ccultivating Bacillus subtilis ER2566 at 37°C carrying the backbone pTXB1 containing bacteriocin gene to produce our target protein as a biostimulator.
- Sample
- Fresh colony of Bacillus subtilis ER2566 carrying the backbone pTXB1 containing Bovicin HJ50 gene from an overnight plate
- Fresh colony of Bacillus subtilis ER2566 carrying the backbone pTXB1 containing Durancin gene from an overnight plate
IPTG Induction
- Inducting Bacillus subtilis ER2566 carrying the backbone pTXB1 containing bacteriocin gene after cultivating at 37°C to produce our target protein as a biostimulator.
- Sample
- Culture of Bacillus subtilis carrying the backbone pTXB1 containing Bovicin HJ50 gene
- Culture of Bacillus subtilis carrying the backbone pTXB1 containing Durancin gene
August 25
Sonication
- Breaking Bacillus subtilis ER2566 carrying the backbone pTXB1 containing bacteriocin gene after cultivating to harvest the protein by sonication.
- Sample
- Culture of Bacillus subtilis carrying the backbone pTXB1 containing Bovicin HJ50 gene induced by IPTG
- Culture of Bacillus subtilis carrying the backbone pTXB1 containing Durancin 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 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
September 3
None
September 4
None
September 5
None
September 6
Transformation
- Transforming backbone pTXB1 containing Lacticin Z gene into Bacillus subtilis ER2566 to produce our target protein as a biostimulator.
- Sample
- backbone pTXB1 containing Lacticin Z gene mini
Cultivation
- Cultivate the E. coli ER2566 at 37°Ccultivating Bacillus subtilis ER2566 at 37°C carrying the backbone pTXB1 containing Lacticin Z gene to produce our target protein as a biostimulator.
- Sample
- Fresh colony of Bacillus subtilis ER2566 carrying the backbone pTXB1 containing Lacticin Z gene from an overnight plate
IPTG Induction
- Inducting Bacillus subtilis ER2566 carrying the backbone pTXB1 containing Lacticin Z gene after cultivating at 37°C to produce our target protein as a biostimulator.
- Sample
- Culture of Bacillus subtilis carrying the backbone pTXB1 containing Lacticin Z gene
September 7
Sonication
- Breaking Bacillus subtilis ER2566 carrying the backbone pTXB1 containing Lacticin Z gene after cultivating to harvest the protein by sonication.
- Sample
- Culture of Bacillus subtilis 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
None
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 Bacillus subtilis Rosetta Gami to produce protein as our negative control in verifying the function of our target peptide.
- Transforming backbone pTXB1 containing bacteriocin gene into Bacillus subtilis Rosetta-gami to produce our target protein as a biostimulator.
- 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 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°Ccultivating Bacillus subtilis 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°Ccultivating Bacillus subtilis Rosetta-gami at 37°C carrying the backbone pTXB1 containing bacteriocin gene produce our target protein as a biostimulator.
- Sample
- Fresh colony of Bacillus subtilis Rosetta Gami carrying the backbone pTXB1 from an overnight plate
- Fresh colony of Bacillus subtilis Rosetta Gami carrying the backbone pTXB1 containing Enterocin B gene from an overnight plate
- Fresh colony of Bacillus subtilis Rosetta Gami carrying the backbone pTXB1 containing Enterocin 96 gene from an overnight plate
- Fresh colony of Bacillus subtilis Rosetta Gami carrying the backbone pTXB1 containing Leucocyclicin Q gene from an overnight plate
IPTG Induction
- Cultivate the E. coli ER2566 at 37°Ccultivating Bacillus subtilis 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 Bacillus subtilis Rosetta-gami carrying the backbone pTXB1 containing bacteriocin gene after cultivating at 37°C produce our target protein as a biostimulator.
- Sample
- Fresh colony of Bacillus subtilis Rosetta Gami carrying the backbone pTXB1 from an overnight plate
- Culture of Bacillus subtilis Rosetta Gami carrying the backbone pTXB1 containing Enterocin B gene
- Culture of Bacillus subtilis Rosetta Gami carrying the backbone pTXB1 containing Enterocin 96 gene
- Culture of Bacillus subtilis Rosetta Gami carrying the backbone pTXB1 containing Leucocyclicin Q gene
October 7
Cultivation
- Cultivate the E. coli ER2566 at 37°Ccultivating Bacillus subtilis Rosetta-gami at 37°C carrying the backbone pTXB1 containing bacteriocin gene produce our target protein as a biostimulator.
- Sample
- Fresh colony of Bacillus subtilis Rosetta Gami carrying the backbone pTXB1 containing Bovicin HJ50 gene from an overnight plate
- Fresh colony of Bacillus subtilis Rosetta Gami carrying the backbone pTXB1 containing Durancin gene from an overnight plate
- Fresh colony of Bacillus subtilis Rosetta Gami carrying the backbone pTXB1 containing Lacticin Z gene from an overnight plate
IPTG Induction
- Inducting Bacillus subtilis Rosetta-gami carrying the backbone pTXB1 containing bacteriocin gene after cultivating at 37°C produce our target protein as a biostimulator.
- Sample
- Culture of Bacillus subtilis Rosetta Gami carrying the backbone pTXB1 containing Bovicin HJ50 gene
- Culture of Bacillus subtilis Rosetta Gami carrying the backbone pTXB1 containing Durancin gene
- Culture of Bacillus subtilis Rosetta Gami carrying the backbone pTXB1 containing Lacticin Z gene
Sonication
- Breaking Bacillus subtilis Rosetta Gami carrying the backbone pTXB1 and pTXB1 containing bacteriocin gene after cultivating to harvest the protein by sonication.
- Sample
- Culture of Bacillus subtilis carrying the backbone pTXB1 induced by IPTG
- Culture of Bacillus subtilis carrying the backbone pTXB1 containing Enterocin B gene induced by IPTG
- Culture of Bacillus subtilis carrying the backbone pTXB1 containing Enterocin 96 gene induced by IPTG
- Culture of Bacillus subtilis 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 Bacillus subtilis Rosetta Gami carrying the backbone pTXB1 containing bacteriocin gene after cultivating to harvest the protein by sonication.
- Sample
- Culture of Bacillus subtilis carrying the backbone pTXB1 containing Bovicin HJ50 gene induced by IPTG
- Culture of Bacillus subtilis carrying the backbone pTXB1 containing Durancin gene induced by IPTG
- Culture of Bacillus subtilis carrying the backbone pTXB1 containing Lacticin Z gene induced by IPTG
- Culture of Bacillus subtilis 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 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
None
October 10
None
October 11
None
October 12
None
October 13
None
October 14
None
October 15
None
October 16
None
October 17
None
October 18
None
October 19
None
