| 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"
| Line 713: | Line 713: | ||
<li class="list">Sample</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">Enterocin B</li> | <li class="list">Enterocin B</li> | ||
<li class="list">Enterocin 96</li> | <li class="list">Enterocin 96</li> | ||
| Line 720: | Line 719: | ||
<li class="list">Lacticin Z</li> | <li class="list">Lacticin Z</li> | ||
<li class="list">Leucocyclicin Q</li> | <li class="list">Leucocyclicin Q</li> | ||
| − | |||
</ul> | </ul> | ||
</ul> | </ul> | ||
| Line 732: | Line 730: | ||
<p class="note-title">Growth curve exp</p> | <p class="note-title">Growth curve exp</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">Observe the growth population of | + | <li class="list">Observe the growth population of <i>Bacillus subtilis</i> in different temperature Condition: 37°C, 32°C, 27°C</li> |
</ul> | </ul> | ||
</article> | </article> | ||
| Line 757: | Line 755: | ||
<p class="note-title">Growth curve exp</p> | <p class="note-title">Growth curve exp</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">Observe the growth population of | + | <li class="list">Observe the growth population of <i><i>Bacillus subtilis</i></i> in different pH Condition: pH = 4, 5, 6, 7, 8</li> |
</ul> | </ul> | ||
</article> | </article> | ||
| Line 795: | Line 793: | ||
<p class="note-title">Expression</p> | <p class="note-title">Expression</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">repared competent cell E. coli ER2566</li> | + | <li class="list">repared competent cell <i>E. coli</i> ER2566</li> |
</ul> | </ul> | ||
<p class="note-title">Growth curve exp</p> | <p class="note-title">Growth curve exp</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">Observe the growth population of | + | <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> |
| − | + | ||
</ul> | </ul> | ||
</article> | </article> | ||
| Line 836: | Line 833: | ||
<p class="note-title">Cloning</p> | <p class="note-title">Cloning</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 ligation products (Transform into E. coli DH5α)</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">Taq PCR (PCR of ligation products to amplify insert gene)</li> | ||
<li class="list">Electrophoresis (To check PCR products)</li> | <li class="list">Electrophoresis (To check PCR products)</li> | ||
| Line 865: | Line 862: | ||
<p class="note-title">Cloning</p> | <p class="note-title">Cloning</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"> | + | <li class="list">Sequencing plasmid</li> |
<li class="list">Sample</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;"> | ||
| Line 876: | Line 873: | ||
<article id="July-21" class="note-item"> | <article id="July-21" class="note-item"> | ||
<h2>July 21</h2><hr> | <h2>July 21</h2><hr> | ||
| − | <p class="note-title"> | + | <p class="note-title">None</p> |
</article> | </article> | ||
| Line 898: | Line 895: | ||
<p class="note-title">Cloning</p> | <p class="note-title">Cloning</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 ligation products (Transform into E. coli DH5α)</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">Taq PCR (PCR of ligation products to amplify insert gene)</li> | ||
<li class="list">Electrophoresis (To check PCR products)</li> | <li class="list">Electrophoresis (To check PCR products)</li> | ||
| Line 952: | Line 949: | ||
<p class="note-title">Expression</p> | <p class="note-title">Expression</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 (Transform correct plasmid into E. coli ER2566)</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;"> | <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | ||
<li class="list">Leucocyclicin Q + pTXB1</li> | <li class="list">Leucocyclicin Q + pTXB1</li> | ||
| Line 994: | Line 991: | ||
<p class="note-title">Expression</p> | <p class="note-title">Expression</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"> | + | <li class="list">Transformation (Transform correct plasmid into <i>E. coli</i> ER2566)</li> |
<li class="list">sample</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;"> | ||
| Line 1,007: | Line 1,004: | ||
<p class="note-title">Expression</p> | <p class="note-title">Expression</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"> | + | <li class="list">Cultivation (Cultivation of <i>E. coli</i> ER2566 colonies for IPTG induction)</li> |
| − | <li class="list"> | + | <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> | <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;"> | ||
| Line 1,019: | Line 1,016: | ||
<p class="note-title">Expression</p> | <p class="note-title">Expression</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"> | + | <li class="list">SDS-PAGE (To check the protein production after induction)</li> |
<li class="list">sample</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;"> | ||
| Line 1,042: | Line 1,039: | ||
<p class="note-title">Cloning</p> | <p class="note-title">Cloning</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"> | + | <li class="list">Amplify the insert gene (Pfu PCR)</li> |
| − | <li class="list"> | + | <li class="list">Electrophoresis (To check PCR products)</li> |
<li class="list">sample</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">Enterocin B</li> | <li class="list">Enterocin B</li> | ||
<li class="list">Enterocin 96 </li> | <li class="list">Enterocin 96 </li> | ||
<li class="list">Duracin TW49M </li> | <li class="list">Duracin TW49M </li> | ||
| − | |||
</ul> | </ul> | ||
</ul> | </ul> | ||
| Line 1,058: | Line 1,053: | ||
<p class="note-title">Cloning</p> | <p class="note-title">Cloning</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"> | + | <li class="list">Digestion of PCR products (Including insert and backbone pTXB1)</li> |
| − | <li class="list"> | + | <li class="list">Ligation of digestion products (Ligase the DNA fragment into backbone pTXB1)</li> |
| − | <li class="list"> | + | <li class="list">Transformation of ligation products (Transform into <i>E. coli</i> DH5α)</li> |
<li class="list">sample</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">Enterocin B + pTXB1</li> | <li class="list">Enterocin B + pTXB1</li> | ||
<li class="list">Enterocin 96 + pTXB1</li> | <li class="list">Enterocin 96 + pTXB1</li> | ||
<li class="list">Duracin TW49M + pTXB1</li> | <li class="list">Duracin TW49M + pTXB1</li> | ||
| − | |||
</ul> | </ul> | ||
</ul> | </ul> | ||
<p class="note-title">Growth curve</p> | <p class="note-title">Growth curve</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"> | + | <li class="list">Observe the growth of <i><i>Bacillus subtilis</i></i> in different temperature, pH, salinityCondition:</li> |
<li class="list">1.Temp: 37°C , pH:7 , salinity: 0.17M</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">2.Temp: 30°C , pH:9 , salinity: 0.5M</li> | ||
| Line 1,082: | Line 1,075: | ||
<p class="note-title">Cloning</p> | <p class="note-title">Cloning</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"> | + | <li class="list">Taq PCR (PCR of ligation products to amplify insert gene)</li> |
| − | <li class="list"> | + | <li class="list">Electrophoresis (To check PCR products)</li> |
<li class="list">sample</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">Enterocin B + pTXB1</li> | <li class="list">Enterocin B + pTXB1</li> | ||
<li class="list">Enterocin 96 + pTXB1</li> | <li class="list">Enterocin 96 + pTXB1</li> | ||
<li class="list">Duracin TW49M + pTXB1</li> | <li class="list">Duracin TW49M + pTXB1</li> | ||
| − | <li class="list"> | + | </ul> |
| + | </ul> | ||
| + | <p class="note-title">Cloning gene of Curcumin biosensor</p> | ||
| + | <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | ||
| + | <li class="list">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">gene segment of αS1-casein</li> | ||
| + | <li class="list">GS linker and T7 promoter</li> | ||
</ul> | </ul> | ||
</ul> | </ul> | ||
| Line 1,104: | Line 1,105: | ||
<li class="list">Sample</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">Enterocin B + pTXB1</li> | <li class="list">Enterocin B + pTXB1</li> | ||
<li class="list">Enterocin 96 + pTXB1</li> | <li class="list">Enterocin 96 + pTXB1</li> | ||
<li class="list">Duracin TW49M + pTXB1</li> | <li class="list">Duracin TW49M + pTXB1</li> | ||
| − | |||
</ul> | </ul> | ||
</ul> | </ul> | ||
<p class="note-title">Expression</p> | <p class="note-title">Expression</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 (Transform correct plasmid into E. coli ER2566)to test different cultivation temperature</li> | + | <li class="list">Transformation (Transform correct plasmid into <i>E. coli</i> ER2566)to test different cultivation temperature</li> |
</ul> | </ul> | ||
</article> | </article> | ||
| Line 1,121: | Line 1,120: | ||
<p class="note-title">Cloning</p> | <p class="note-title">Cloning</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"> | + | <li class="list">Sequencing plasmid</li> |
<li class="list">Sample</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">Enterocin B + pTXB1</li> | <li class="list">Enterocin B + pTXB1</li> | ||
<li class="list">Enterocin 96 + pTXB1</li> | <li class="list">Enterocin 96 + pTXB1</li> | ||
<li class="list">Duracin TW49M + pTXB1</li> | <li class="list">Duracin TW49M + pTXB1</li> | ||
| − | |||
</ul> | </ul> | ||
</ul> | </ul> | ||
| Line 1,134: | Line 1,131: | ||
<p class="note-title">Expression</p> | <p class="note-title">Expression</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"> | + | <li class="list">Cultivation (Cultivation of <i>E. coli</i> ER2566 colonies for IPTG induction)</li> |
| − | <li class="list"> | + | <li class="list">IPTG Induction (To test the temperature of cultivation after induction)Condition: temp = 37, 30, 13.5°C</li> |
| + | </ul> | ||
| + | <p class="note-title">Expression</p> | ||
| + | <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | ||
| + | <li class="list">Transforming the ligation product of the Curcumin biosensor into <i>E. coli</i> BL21 DE3 Cultivating the culture and inducing it with IPTG.</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> BL21 DE3 carrying the backbone pet30a containing αS1-casei and GS linker gene from an overnight plate</li> | ||
</ul> | </ul> | ||
</article> | </article> | ||
| Line 1,143: | Line 1,147: | ||
<p class="note-title">Expression</p> | <p class="note-title">Expression</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"> | + | <li class="list">SDS-PAGE (To check the protein production after induction)Check different temperature</li> |
| − | <li class="list"> | + | <li class="list">IPTG Induction (To test the concentration of IPTG)Condition: concentration = 200, 400, 600, 800, 1000μM</li> |
</ul> | </ul> | ||
</article> | </article> | ||
| Line 1,152: | Line 1,156: | ||
<p class="note-title">Expression</p> | <p class="note-title">Expression</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"> | + | <li class="list">SDS-PAGE (To check the protein production after induction) -> Check different IPTG concentration</li> |
| + | </ul> | ||
| + | <p class="note-title">Chip Production</p> | ||
| + | <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | ||
| + | <li class="list">Processing the chips with Mua , EtOH , and EDC+NHS mixture, then adding αS1-casein on chips.</li> | ||
<li class="list">Sample</li> | <li class="list">Sample</li> | ||
| + | <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | ||
| + | <li class="list">αS1-casein protein</li> | ||
| + | |||
</ul> | </ul> | ||
</article> | </article> | ||
| Line 1,159: | Line 1,170: | ||
<article id="August-18" class="note-item"> | <article id="August-18" class="note-item"> | ||
<h2>August 18</h2><hr> | <h2>August 18</h2><hr> | ||
| − | <p class="note-title"> | + | <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><i>Bacillus subtilis</i></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><i>Bacillus subtilis</i></i> ER2566 to produce our target protein as a biostimulator.</li> | ||
| + | <li class="list">Sample</li> | ||
| + | <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | ||
| + | <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 Leucocyclicin Q gene mini</li> | ||
| + | </ul> | ||
| + | <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°Ccultivating <i><i>Bacillus subtilis</i></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°Ccultivating <i><i>Bacillus subtilis</i></i> ER2566 at 37°C carrying the backbone ptxB1 containing bacteriocin gene to produce our target protein as a biostimulator.</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><i>Bacillus subtilis</i></i> ER2566 carrying the backbone ptxB1 from an overnight plate</li> | ||
| + | <li class="list">Fresh colony of <i><i>Bacillus subtilis</i></i> ER2566 carrying the backbone ptxB1 containing Enterocin B gene from an overnight plate</li> | ||
| + | <li class="list">Fresh colony of <i><i>Bacillus subtilis</i></i> ER2566 carrying the backbone ptxB1 containing Enterocin 96 gene from an overnight plate</li> | ||
| + | <li class="list">Fresh colony of <i><i>Bacillus subtilis</i></i> ER2566 carrying the backbone ptxB1 containing Leucocyclicin Q gene from an overnight plate</li> | ||
| + | </ul> | ||
| + | <p class="note-title">IPTG Induction</p> | ||
| + | <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | ||
| + | <li class="list">Inducting <i><i>Bacillus subtilis</i></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">Inducting <i><i>Bacillus subtilis</i></i> ER2566 carrying the backbone ptxB1 containing bacteriocin gene after cultivating at 37°C to produce our target protein as a biostimulator.</li> | ||
| + | <li class="list">Sample</li> | ||
| + | <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | ||
| + | <li class="list">Culture of <i><i>Bacillus subtilis</i></i> carrying the backbone ptxB1</li> | ||
| + | <li class="list">Culture of <i><i>Bacillus subtilis</i></i> carrying the backbone ptxB1 containing Enterocin B gene</li> | ||
| + | <li class="list">Culture of <i><i>Bacillus subtilis</i></i> carrying the backbone ptxB1 containing Enterocin 96 gene</li> | ||
| + | <li class="list">Culture of <i><i>Bacillus subtilis</i></i> carrying the backbone ptxB1 containing Leucocyclicin Q gene</li> | ||
| + | </ul> | ||
</article> | </article> | ||
| Line 1,166: | Line 1,209: | ||
<p class="note-title">Cloning</p> | <p class="note-title">Cloning</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"> | + | <li class="list">Digestion of PCR products (Including insert and backbone pET30a)</li> |
| − | <li class="list"> | + | <li class="list">Ligation of digestion products (Ligase the DNA fragment into backbone pET30a)</li> |
<li class="list">Sample(8/17)</li> | <li class="list">Sample(8/17)</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">Enterocin B + pET30a</li> | <li class="list">Enterocin B + pET30a</li> | ||
<li class="list">Enterocin 96 + pET30a</li> | <li class="list">Enterocin 96 + pET30a</li> | ||
| Line 1,177: | Line 1,219: | ||
<li class="list">Lacticin Z + pET30a</li> | <li class="list">Lacticin Z + pET30a</li> | ||
<li class="list">Leucocyclicin Q + pET30a</li> | <li class="list">Leucocyclicin Q + pET30a</li> | ||
| − | |||
</ul> | </ul> | ||
</ul> | </ul> | ||
| + | <p class="note-title">Sonication</p> | ||
| + | <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | ||
| + | <li class="list">Breaking <i><i>Bacillus subtilis</i></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">Culture of <i><i>Bacillus subtilis</i></i> carrying the backbone ptxB1 induced by IPTG</li> | ||
| + | <li class="list">Culture of <i><i>Bacillus subtilis</i></i> carrying the backbone ptxB1 containing Enterocin B gene induced by IPTG </li> | ||
| + | <li class="list">Culture of <i><i>Bacillus subtilis</i></i> carrying the backbone ptxB1 containing Enterocin 96 gene induced by IPTG </li> | ||
| + | <li class="list">Culture of <i><i>Bacillus subtilis</i></i> carrying the backbone ptxB1 containing Leucocyclicin Q gene induced by IPTG </li> | ||
| + | </ul> | ||
| + | </ul> | ||
| + | <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">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">Protein expressing the backbone ptxB1 induced by IPTG</li> | ||
| + | <li class="list">Protein expressing the backbone ptxB1 containing Enterocin B gene induced by IPTG </li> | ||
| + | <li class="list">Protein expressing the backbone ptxB1 containing Enterocin 96 gene induced by IPTG </li> | ||
| + | <li class="list">Protein expressing the backbone ptxB1 containing Leucocyclicin Q gene induced by IPTG </li> | ||
| + | </ul> | ||
| + | </ul> | ||
| + | <p class="note-title">Biosensor Measurement 1- Electrochemical Impedance Spectroscopy</p> | ||
</article> | </article> | ||
| Line 1,186: | Line 1,250: | ||
<p class="note-title">Cloning</p> | <p class="note-title">Cloning</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"> | + | <li class="list">Transformation of ligation products (Transform into <i>E. coli</i> DH5α)</li> |
| − | <li class="list"> | + | <li class="list">Taq PCR (PCR of ligation products to amplify insert gene)</li> |
| − | <li class="list"> | + | <li class="list">Electrophoresis (To check PCR products)</li> |
<li class="list">Sample</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">Enterocin B + pET30a</li> | <li class="list">Enterocin B + pET30a</li> | ||
<li class="list">Enterocin 96 + pET30a</li> | <li class="list">Enterocin 96 + pET30a</li> | ||
| Line 1,198: | Line 1,261: | ||
<li class="list">Lacticin Z + pET30a</li> | <li class="list">Lacticin Z + pET30a</li> | ||
<li class="list">Leucocyclicin Q + pET30a</li> | <li class="list">Leucocyclicin Q + pET30a</li> | ||
| − | |||
</ul> | </ul> | ||
</ul> | </ul> | ||
| Line 1,207: | Line 1,269: | ||
<p class="note-title">Cloning</p> | <p class="note-title">Cloning</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"> | + | <li class="list">Cultivation</li> |
| − | <li class="list"> | + | <li class="list">Miniprep (Purify Plasmid)</li> |
<li class="list">Sample</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">Enterocin B + pET30a</li> | <li class="list">Enterocin B + pET30a</li> | ||
<li class="list">Enterocin 96 + pET30a</li> | <li class="list">Enterocin 96 + pET30a</li> | ||
| Line 1,218: | Line 1,279: | ||
<li class="list">Lacticin Z + pET30a</li> | <li class="list">Lacticin Z + pET30a</li> | ||
<li class="list">Leucocyclicin Q + pET30a</li> | <li class="list">Leucocyclicin Q + pET30a</li> | ||
| − | |||
</ul> | </ul> | ||
</article> | </article> | ||
| Line 1,229: | Line 1,289: | ||
<li class="list">Sample</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">Enterocin B + pET30a</li> | <li class="list">Enterocin B + pET30a</li> | ||
<li class="list">Enterocin 96 + pET30a</li> | <li class="list">Enterocin 96 + pET30a</li> | ||
| Line 1,236: | Line 1,295: | ||
<li class="list">Lacticin Z + pET30a</li> | <li class="list">Lacticin Z + pET30a</li> | ||
<li class="list">Leucocyclicin Q + pET30a</li> | <li class="list">Leucocyclicin Q + pET30a</li> | ||
| − | |||
</ul> | </ul> | ||
</ul> | </ul> | ||
| + | <p class="note-title">Biosensor Measurement 2- Differential Pulse Voltammetry</p> | ||
</article> | </article> | ||
<article id="August-23" class="note-item"> | <article id="August-23" class="note-item"> | ||
<h2>August 23</h2><hr> | <h2>August 23</h2><hr> | ||
| − | <p class="note-title"> | + | <p class="note-title">None</p> |
| − | + | ||
| − | + | ||
| − | + | ||
| − | + | ||
| − | + | ||
| − | + | ||
| − | + | ||
| − | + | ||
| − | + | ||
| − | + | ||
| − | + | ||
| − | + | ||
| − | + | ||
| − | + | ||
| − | + | ||
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| − | + | ||
| − | + | ||
| − | + | ||
| − | + | ||
| − | + | ||
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| − | + | ||
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| − | + | ||
| − | + | ||
| − | + | ||
</article> | </article> | ||
<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><i>Bacillus subtilis</i></i> ER2566 to produce our target protein as a biostimulator.</li> | ||
<li class="list">Sample</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"> | + | <li class="list">backbone ptxB1 containing Bovicin HJ50 gene mini</li> |
| + | <li class="list">backbone ptxB1 containing Durancin gene mini</li> | ||
</ul> | </ul> | ||
| − | + | <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;"> | ||
| − | <li class="list"> | + | <li class="list">Cultivate the <i>E. coli</i> ER2566 at 37°Ccultivating <i><i>Bacillus subtilis</i></i> ER2566 at 37°C carrying the backbone ptxB1 containing bacteriocin gene to produce our target protein as a biostimulator.</li> |
<li class="list">Sample</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"> | + | <li class="list">Fresh colony of <i><i>Bacillus subtilis</i></i> ER2566 carrying the backbone ptxB1 containing Bovicin HJ50 gene from an overnight plate</li> |
| − | + | <li class="list">Fresh colony of <i><i>Bacillus subtilis</i></i> ER2566 carrying the backbone ptxB1 containing Durancin gene from an overnight plate</li> | |
| − | + | </ul> | |
| − | + | <p class="note-title">IPTG Induction</p> | |
| − | + | ||
| − | + | ||
| − | + | ||
| − | + | ||
| − | </ul> | + | |
| − | + | ||
| − | + | ||
| − | + | ||
| − | + | ||
<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">Inducting <i><i>Bacillus subtilis</i></i> ER2566 carrying the backbone ptxB1 containing bacteriocin gene after cultivating at 37°C to produce our target protein as a biostimulator.</li> | |
<li class="list">Sample</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"> | + | <li class="list">Culture of <i><i>Bacillus subtilis</i></i> carrying the backbone ptxB1 containing Bovicin HJ50 gene</li> |
| − | <li class="list"> | + | <li class="list">Culture of <i><i>Bacillus subtilis</i></i> carrying the backbone ptxB1 containing Durancin gene</li> |
</ul> | </ul> | ||
| − | |||
| − | |||
| − | |||
| − | |||
| − | |||
| − | |||
| − | |||
| − | |||
| − | |||
| − | |||
</article> | </article> | ||
<article id="August-25" class="note-item"> | <article id="August-25" class="note-item"> | ||
<h2>August 25</h2><hr> | <h2>August 25</h2><hr> | ||
| − | + | <p class="note-title">Sonication</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">Breaking <i><i>Bacillus subtilis</i></i> ER2566 carrying the backbone ptxB1 containing bacteriocin gene after cultivating to harvest the protein by sonication.</li> | ||
<li class="list">Sample</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"> | + | <li class="list">Culture of <i><i>Bacillus subtilis</i></i> carrying the backbone ptxB1 containing Bovicin HJ50 gene induced by IPTG </li> |
| + | <li class="list">Culture of <i><i>Bacillus subtilis</i></i> carrying the backbone ptxB1 containing Durancin gene induced by IPTG </li> | ||
</ul> | </ul> | ||
| − | |||
</ul> | </ul> | ||
| − | + | <p class="note-title">Protein quantification & SDS-PAGE</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"> | + | <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> | <li class="list">Sample</li> | ||
| − | + | <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | |
| − | <li class="list"> | + | <li class="list">Protein expressing the backbone ptxB1 containing Bovicin HJ50 gene induced by IPTG </li> |
| − | + | <li class="list">Protein expressing the backbone ptxB1 containing Durancin gene induced by IPTG </li> | |
| + | </ul> | ||
</ul> | </ul> | ||
</article> | </article> | ||
| Line 1,345: | Line 1,357: | ||
<article id="August-26" class="note-item"> | <article id="August-26" class="note-item"> | ||
<h2>August 26</h2><hr> | <h2>August 26</h2><hr> | ||
| − | <p class="note-title"> | + | <p class="note-title">None</p> |
| − | + | ||
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</article> | </article> | ||
<article id="August-27" class="note-item"> | <article id="August-27" class="note-item"> | ||
<h2>August 27</h2><hr> | <h2>August 27</h2><hr> | ||
| − | <p class="note-title"> | + | <p class="note-title">None</p> |
| − | + | ||
| − | + | ||
| − | + | ||
| − | + | ||
| − | + | ||
| − | + | ||
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| − | + | ||
</article> | </article> | ||
| + | |||
<article id="August-28" class="note-item"> | <article id="August-28" class="note-item"> | ||
<h2>August 28</h2><hr> | <h2>August 28</h2><hr> | ||
| − | <p class="note-title"> | + | <p class="note-title">None</p> |
| − | + | ||
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</article> | </article> | ||
<article id="August-29" class="note-item"> | <article id="August-29" class="note-item"> | ||
<h2>August 29</h2><hr> | <h2>August 29</h2><hr> | ||
| − | <p class="note-title"> | + | <p class="note-title">None</p> |
| − | + | ||
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</article> | </article> | ||
<article id="August-30" class="note-item"> | <article id="August-30" class="note-item"> | ||
<h2>August 30</h2><hr> | <h2>August 30</h2><hr> | ||
| − | <p class="note-title"> | + | <p class="note-title">None</p> |
| − | + | ||
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</article> | </article> | ||
<article id="August-31" class="note-item"> | <article id="August-31" class="note-item"> | ||
<h2>August 31</h2><hr> | <h2>August 31</h2><hr> | ||
| − | <p class="note-title"> | + | <p class="note-title">None</p> |
| − | + | ||
| − | + | ||
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</article> | </article> | ||
| Line 1,756: | Line 1,389: | ||
<article id="September-1" class="note-item"> | <article id="September-1" class="note-item"> | ||
<h2>September 1</h2><hr> | <h2>September 1</h2><hr> | ||
| − | <p class="note-title"> | + | <p class="note-title">None</p> |
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</article> | </article> | ||
<article id="September-2" class="note-item"> | <article id="September-2" class="note-item"> | ||
<h2>September 2</h2><hr> | <h2>September 2</h2><hr> | ||
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</article> | </article> | ||
<article id="September-3" class="note-item"> | <article id="September-3" class="note-item"> | ||
<h2>September 3</h2><hr> | <h2>September 3</h2><hr> | ||
| − | <p class="note-title"> | + | <p class="note-title">None</p> |
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</article> | </article> | ||
<article id="September-4" class="note-item"> | <article id="September-4" class="note-item"> | ||
<h2>September 4</h2><hr> | <h2>September 4</h2><hr> | ||
| − | <p class="note-title"> | + | <p class="note-title">None</p> |
| − | + | ||
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</article> | </article> | ||
<article id="September-5" class="note-item"> | <article id="September-5" class="note-item"> | ||
<h2>September 5</h2><hr> | <h2>September 5</h2><hr> | ||
| − | <p class="note-title"> | + | <p class="note-title">None</p> |
| − | + | </article> | |
| − | + | ||
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| − | + | <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;"> | <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><i>Bacillus subtilis</i></i> ER2566 to produce our target protein as a biostimulator.</li> | ||
<li class="list">Sample</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"> | + | <li class="list">backbone ptxB1 containing Lacticin Z gene mini</li> |
| − | + | ||
| − | + | ||
| − | + | ||
</ul> | </ul> | ||
| − | + | <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;"> | ||
| − | <li class="list"> | + | <li class="list">Cultivate the <i>E. coli</i> ER2566 at 37°Ccultivating <i><i>Bacillus subtilis</i></i> ER2566 at 37°C carrying the backbone ptxB1 containing Lacticin Z gene to produce our target protein as a biostimulator.</li> |
<li class="list">Sample</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"> | + | <li class="list">Fresh colony of <i><i>Bacillus subtilis</i></i> ER2566 carrying the backbone ptxB1 containing Lacticin Z gene from an overnight plate</li> |
| − | + | </ul> | |
| − | + | <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;"> | ||
| − | <li class="list"> | + | <li class="list">Inducting <i><i>Bacillus subtilis</i></i> ER2566 carrying the backbone ptxB1 containing Lacticin Z gene after cultivating at 37°C to produce our target protein as a biostimulator.</li> |
<li class="list">Sample</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"> | + | <li class="list">Culture of <i><i>Bacillus subtilis</i></i> carrying the backbone ptxB1 containing Lacticin Z gene</li> |
| − | + | ||
| − | + | ||
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</ul> | </ul> | ||
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</article> | </article> | ||
<article id="September-7" class="note-item"> | <article id="September-7" class="note-item"> | ||
<h2>September 7</h2><hr> | <h2>September 7</h2><hr> | ||
| − | + | <p class="note-title">Sonication</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"> | + | <li class="list">Breaking <i><i>Bacillus subtilis</i></i> ER2566 carrying the backbone ptxB1 containing Lacticin Z gene after cultivating to harvest the protein by sonication.</li> |
<li class="list">Sample</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">Culture of <i><i>Bacillus subtilis</i></i> carrying the backbone ptxB1 containing Lacticin Z gene induced by IPTG </li> | |
| − | + | ||
</ul> | </ul> | ||
| − | |||
</ul> | </ul> | ||
| − | + | <p class="note-title">Protein quantification & SDS-PAGE</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"> | + | <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> | <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">Protein expressing the backbone ptxB1 containing Lacticin Z gene induced by IPTG </li> | |
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</ul> | </ul> | ||
</ul> | </ul> | ||
| Line 2,111: | Line 1,458: | ||
<article id="September-8" class="note-item"> | <article id="September-8" class="note-item"> | ||
<h2>September 8</h2><hr> | <h2>September 8</h2><hr> | ||
| − | <p class="note-title"> | + | <p class="note-title">None</p> |
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</article> | </article> | ||
<article id="September-9" class="note-item"> | <article id="September-9" class="note-item"> | ||
<h2>September 9</h2><hr> | <h2>September 9</h2><hr> | ||
| − | <p class="note-title"> | + | <p class="note-title">None</p> |
| − | + | ||
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</article> | </article> | ||
| Line 2,248: | Line 1,473: | ||
<p class="note-title">None</p> | <p class="note-title">None</p> | ||
</article> | </article> | ||
| + | |||
<article id="September-11" class="note-item"> | <article id="September-11" class="note-item"> | ||
<h2>September 11</h2><hr> | <h2>September 11</h2><hr> | ||
| − | <p class="note-title"> | + | <p class="note-title">None</p> |
| − | + | ||
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</article> | </article> | ||
<article id="September-12" class="note-item"> | <article id="September-12" class="note-item"> | ||
<h2>September 12</h2><hr> | <h2>September 12</h2><hr> | ||
| − | <p class="note-title"> | + | <p class="note-title">None</p> |
| − | + | ||
| − | + | ||
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</article> | </article> | ||
| Line 2,285: | Line 1,488: | ||
<p class="note-title">None</p> | <p class="note-title">None</p> | ||
</article> | </article> | ||
| + | |||
<article id="September-14" class="note-item"> | <article id="September-14" class="note-item"> | ||
<h2>September 14</h2><hr> | <h2>September 14</h2><hr> | ||
<p class="note-title">None</p> | <p class="note-title">None</p> | ||
</article> | </article> | ||
| + | |||
<article id="September-15" class="note-item"> | <article id="September-15" class="note-item"> | ||
<h2>September 15</h2><hr> | <h2>September 15</h2><hr> | ||
<p class="note-title">None</p> | <p class="note-title">None</p> | ||
</article> | </article> | ||
| + | |||
<article id="September-16" class="note-item"> | <article id="September-16" class="note-item"> | ||
<h2>September 16</h2><hr> | <h2>September 16</h2><hr> | ||
<p class="note-title">None</p> | <p class="note-title">None</p> | ||
</article> | </article> | ||
| + | |||
<article id="September-17" class="note-item"> | <article id="September-17" class="note-item"> | ||
<h2>September 17</h2><hr> | <h2>September 17</h2><hr> | ||
| − | <p class="note-title"> | + | <p class="note-title">None</p> |
| − | + | ||
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</article> | </article> | ||
| Line 2,318: | Line 1,513: | ||
<p class="note-title">None</p> | <p class="note-title">None</p> | ||
</article> | </article> | ||
| + | |||
<article id="September-19" class="note-item"> | <article id="September-19" class="note-item"> | ||
<h2>September 19</h2><hr> | <h2>September 19</h2><hr> | ||
<p class="note-title">None</p> | <p class="note-title">None</p> | ||
</article> | </article> | ||
| + | |||
<article id="September-20" class="note-item"> | <article id="September-20" class="note-item"> | ||
<h2>September 20</h2><hr> | <h2>September 20</h2><hr> | ||
<p class="note-title">None</p> | <p class="note-title">None</p> | ||
</article> | </article> | ||
| + | |||
<article id="September-21" class="note-item"> | <article id="September-21" class="note-item"> | ||
<h2>September 21</h2><hr> | <h2>September 21</h2><hr> | ||
<p class="note-title">None</p> | <p class="note-title">None</p> | ||
</article> | </article> | ||
| + | |||
<article id="September-22" class="note-item"> | <article id="September-22" class="note-item"> | ||
<h2>September 22</h2><hr> | <h2>September 22</h2><hr> | ||
| − | + | <p class="note-title">None</p> | |
| − | + | ||
| − | + | ||
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</article> | </article> | ||
<article id="September-23" class="note-item"> | <article id="September-23" class="note-item"> | ||
<h2>September 23</h2><hr> | <h2>September 23</h2><hr> | ||
| − | + | <p class="note-title">None</p> | |
| − | + | ||
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</article> | </article> | ||
<article id="September-24" class="note-item"> | <article id="September-24" class="note-item"> | ||
<h2>September 24</h2><hr> | <h2>September 24</h2><hr> | ||
| − | + | <p class="note-title">None</p> | |
| − | + | ||
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</article> | </article> | ||
<article id="September-25" class="note-item"> | <article id="September-25" class="note-item"> | ||
<h2>September 25</h2><hr> | <h2>September 25</h2><hr> | ||
| − | + | <p class="note-title">None</p> | |
| − | + | ||
| − | + | ||
| − | + | ||
| − | + | ||
| − | + | ||
| − | + | ||
| − | + | ||
| − | + | ||
| − | + | ||
| − | + | ||
| − | + | ||
</article> | </article> | ||
| Line 2,430: | Line 1,553: | ||
<p class="note-title">None</p> | <p class="note-title">None</p> | ||
</article> | </article> | ||
| + | |||
<article id="September-27" class="note-item"> | <article id="September-27" class="note-item"> | ||
<h2>September 27</h2><hr> | <h2>September 27</h2><hr> | ||
<p class="note-title">None</p> | <p class="note-title">None</p> | ||
</article> | </article> | ||
| + | |||
<article id="September-28" class="note-item"> | <article id="September-28" class="note-item"> | ||
<h2>September 28</h2><hr> | <h2>September 28</h2><hr> | ||
| − | <p class="note-title"> | + | <p class="note-title">None</p> |
| − | + | ||
| − | + | ||
| − | + | ||
| − | + | ||
| − | + | ||
| − | + | ||
| − | + | ||
| − | + | ||
| − | + | ||
</article> | </article> | ||
| + | |||
<article id="September-29" class="note-item"> | <article id="September-29" class="note-item"> | ||
<h2>September 29</h2><hr> | <h2>September 29</h2><hr> | ||
<p class="note-title">None</p> | <p class="note-title">None</p> | ||
</article> | </article> | ||
| + | |||
<article id="September-30" class="note-item"> | <article id="September-30" class="note-item"> | ||
<h2>September 30</h2><hr> | <h2>September 30</h2><hr> | ||
| Line 2,462: | Line 1,580: | ||
<p class="note-title">None</p> | <p class="note-title">None</p> | ||
</article> | </article> | ||
| + | |||
<article id="October-2" class="note-item"> | <article id="October-2" class="note-item"> | ||
<h2>October 2</h2><hr> | <h2>October 2</h2><hr> | ||
<p class="note-title">None</p> | <p class="note-title">None</p> | ||
</article> | </article> | ||
| + | |||
<article id="October-3" class="note-item"> | <article id="October-3" class="note-item"> | ||
<h2>October 3</h2><hr> | <h2>October 3</h2><hr> | ||
<p class="note-title">None</p> | <p class="note-title">None</p> | ||
</article> | </article> | ||
| + | |||
<article id="October-4" class="note-item"> | <article id="October-4" class="note-item"> | ||
<h2>October 4</h2><hr> | <h2>October 4</h2><hr> | ||
<p class="note-title">None</p> | <p class="note-title">None</p> | ||
</article> | </article> | ||
| + | |||
<article id="October-5" class="note-item"> | <article id="October-5" class="note-item"> | ||
<h2>October 5</h2><hr> | <h2>October 5</h2><hr> | ||
| − | <p class="note-title"> | + | <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"> | + | <li class="list">Transforming backbone ptxB1 into <i><i>Bacillus subtilis</i></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><i>Bacillus subtilis</i></i> Rosetta-gami to produce our target protein as a biostimulator.</li> | ||
<li class="list">Sample</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"> | + | <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 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> | ||
</ul> | </ul> | ||
| Line 2,488: | Line 1,617: | ||
<article id="October-6" class="note-item"> | <article id="October-6" class="note-item"> | ||
<h2>October 6</h2><hr> | <h2>October 6</h2><hr> | ||
| − | <p class="note-title"> | + | <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°Ccultivating <i><i>Bacillus subtilis</i></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°Ccultivating <i><i>Bacillus subtilis</i></i> Rosetta-gami at 37°C carrying the backbone ptxB1 containing bacteriocin gene produce our target protein as a biostimulator.</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><i>Bacillus subtilis</i></i> Rosetta Gami carrying the backbone ptxB1 from an overnight plate</li> | ||
| + | <li class="list">Fresh colony of <i><i>Bacillus subtilis</i></i> Rosetta Gami carrying the backbone ptxB1 containing Enterocin B gene from an overnight plate</li> | ||
| + | <li class="list">Fresh colony of <i><i>Bacillus subtilis</i></i> Rosetta Gami carrying the backbone ptxB1 containing Enterocin 96 gene from an overnight plate</li> | ||
| + | <li class="list">Fresh colony of <i><i>Bacillus subtilis</i></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°Ccultivating <i><i>Bacillus subtilis</i></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><i>Bacillus subtilis</i></i> Rosetta-gami carrying the backbone ptxB1 containing bacteriocin gene after cultivating at 37°C produce our target protein as a biostimulator.</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><i>Bacillus subtilis</i></i> Rosetta Gami carrying the backbone ptxB1 from an overnight plate</li> | ||
| + | <li class="list">Culture of <i><i>Bacillus subtilis</i></i> Rosetta Gami carrying the backbone ptxB1 containing Enterocin B gene</li> | ||
| + | <li class="list">Culture of <i><i>Bacillus subtilis</i></i> Rosetta Gami carrying the backbone ptxB1 containing Enterocin 96 gene</li> | ||
| + | <li class="list">Culture of <i><i>Bacillus subtilis</i></i> Rosetta Gami carrying the backbone ptxB1 containing Leucocyclicin Q gene</li> | ||
| + | </ul> | ||
| + | </ul> | ||
</article> | </article> | ||
<article id="October-7" class="note-item"> | <article id="October-7" class="note-item"> | ||
<h2>October 7</h2><hr> | <h2>October 7</h2><hr> | ||
| − | <p class="note-title"> | + | <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;"> | ||
| + | <li class="list">Cultivate the <i>E. coli</i> ER2566 at 37°Ccultivating <i><i>Bacillus subtilis</i></i> Rosetta-gami at 37°C carrying the backbone ptxB1 containing bacteriocin gene produce our target protein as a biostimulator.</li> | ||
<li class="list">Sample</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"> | + | <li class="list">Fresh colony of <i><i>Bacillus subtilis</i></i> Rosetta Gami carrying the backbone ptxB1 containing Bovicin HJ50 gene from an overnight plate</li> |
| − | + | <li class="list">Fresh colony of <i><i>Bacillus subtilis</i></i> Rosetta Gami carrying the backbone ptxB1 containing Durancin gene from an overnight plate</li> | |
| − | + | <li class="list">Fresh colony of <i><i>Bacillus subtilis</i></i> Rosetta Gami carrying the backbone ptxB1 containing Lacticin Z gene from an overnight plate</li> | |
| − | <li class="list"> | + | |
| − | + | ||
| − | + | ||
| − | <li class="list"> | + | |
| − | + | ||
| − | + | ||
| − | + | ||
</ul> | </ul> | ||
</ul> | </ul> | ||
| − | + | <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;"> | ||
| + | <li class="list">Inducting <i><i>Bacillus subtilis</i></i> Rosetta-gami carrying the backbone ptxB1 containing bacteriocin gene after cultivating at 37°C produce our target protein as a biostimulator.</li> | ||
<li class="list">Sample</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"> | + | <li class="list">Culture of <i><i>Bacillus subtilis</i></i> Rosetta Gami carrying the backbone ptxB1 containing Bovicin HJ50 gene</li> |
| − | + | <li class="list">Culture of <i><i>Bacillus subtilis</i></i> Rosetta Gami carrying the backbone ptxB1 containing Durancin gene</li> | |
| − | + | <li class="list">Culture of <i><i>Bacillus subtilis</i></i> Rosetta Gami carrying the backbone ptxB1 containing Lacticin Z gene</li> | |
| − | <li class="list"> | + | |
| − | + | ||
| − | <li class="list"> | + | |
| − | + | ||
</ul> | </ul> | ||
</ul> | </ul> | ||
| − | + | <p class="note-title">Sonication</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">Breaking <i><i>Bacillus subtilis</i></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> | <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"> | + | <li class="list">Culture of <i><i>Bacillus subtilis</i></i> carrying the backbone ptxB1 induced by IPTG </li> |
| − | + | <li class="list">Culture of <i><i>Bacillus subtilis</i></i> carrying the backbone ptxB1 containing Enterocin B gene induced by IPTG</li> | |
| − | <li class="list"> | + | <li class="list">Culture of <i><i>Bacillus subtilis</i></i> carrying the backbone ptxB1 containing Enterocin 96 gene induced by IPTG</li> |
| − | + | <li class="list">Culture of <i><i>Bacillus subtilis</i></i> carrying the backbone ptxB1 containing Leucocyclicin Q gene induced by IPTG</li> | |
| − | <li class="list"> | + | |
| − | + | ||
| − | <li class="list"> | + | |
| − | + | ||
</ul> | </ul> | ||
</ul> | </ul> | ||
| − | + | <p class="note-title">Protein quantification & SDS-PAGE</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"> | + | <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> | <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"> | + | <li class="list">Protein expressing the backbone ptxB1 induced by IPTG</li> |
| − | <li class="list"> | + | <li class="list">Protein expressing the backbone ptxB1 containing Enterocin B gene induced by IPTG </li> |
| − | <li class="list"> | + | <li class="list">Protein expressing the backbone ptxB1 containing Enterocin 96 gene induced by IPTG </li> |
| − | <li class="list"> | + | <li class="list">Protein expressing the backbone ptxB1 containing Leucocyclicin Q gene induced by IPTG </li> |
| − | + | ||
| − | + | ||
</ul> | </ul> | ||
</ul> | </ul> | ||
| + | </article> | ||
| − | <p class="note-title"> | + | <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;"> | <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | ||
| + | <li class="list">Breaking <i><i>Bacillus subtilis</i></i> Rosetta Gami carrying the backbone ptxB1 containing bacteriocin gene after cultivating to harvest the protein by sonication.</li> | ||
<li class="list">Sample</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"> | + | <li class="list">Culture of <i><i>Bacillus subtilis</i></i> carrying the backbone ptxB1 containing Bovicin HJ50 gene induced by IPTG</li> |
| − | + | <li class="list">Culture of <i><i>Bacillus subtilis</i></i> carrying the backbone ptxB1 containing Durancin gene induced by IPTG</li> | |
| − | <li class="list"> | + | <li class="list">Culture of <i><i>Bacillus subtilis</i></i> carrying the backbone ptxB1 containing Lacticin Z gene induced by IPTG</li> |
| − | + | <li class="list">Culture of <i><i>Bacillus subtilis</i></i> carrying the backbone ptxB1 containing Leucocyclicin Q gene induced by IPTG</li> | |
| − | <li class="list"> | + | |
| − | <li class="list"> | + | |
| − | + | ||
</ul> | </ul> | ||
</ul> | </ul> | ||
| − | + | <p class="note-title">Protein quantification & SDS-PAGE</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">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> | <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"> | + | <li class="list">Protein expressing the backbone ptxB1 containing Bovicin HJ50 gene induced by IPTG </li> |
| − | <li class="list"> | + | <li class="list">Protein expressing the backbone ptxB1 containing Durancin gene induced by IPTG </li> |
| − | <li class="list"> | + | <li class="list">Protein expressing the backbone ptxB1 containing Lacticin Z gene induced by IPTG </li> |
| − | <li class="list"> | + | <li class="list">Protein expressing the backbone ptxB1 containing Leucocyclicin Q gene induced by IPTG </li> |
| − | + | </ul> | |
| − | + | ||
| − | + | ||
| − | + | ||
| − | + | ||
| − | + | ||
| − | + | ||
| − | + | ||
| − | + | ||
| − | + | ||
</ul> | </ul> | ||
| + | </article> | ||
| + | |||
| + | <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">None</p> | ||
</article> | </article> | ||
| Line 2,607: | Line 1,733: | ||
<p class="note-title">None</p> | <p class="note-title">None</p> | ||
</article> | </article> | ||
| + | |||
<article id="October-13" class="note-item"> | <article id="October-13" class="note-item"> | ||
<h2>October 13</h2><hr> | <h2>October 13</h2><hr> | ||
<p class="note-title">None</p> | <p class="note-title">None</p> | ||
</article> | </article> | ||
| + | |||
<article id="October-14" class="note-item"> | <article id="October-14" class="note-item"> | ||
<h2>October 14</h2><hr> | <h2>October 14</h2><hr> | ||
<p class="note-title">None</p> | <p class="note-title">None</p> | ||
</article> | </article> | ||
| + | |||
<article id="October-15" class="note-item"> | <article id="October-15" class="note-item"> | ||
<h2>October 15</h2><hr> | <h2>October 15</h2><hr> | ||
| − | <p class="note-title"> | + | <p class="note-title">None</p> |
| − | + | ||
| − | + | ||
| − | + | ||
| − | + | ||
| − | + | ||
| − | + | ||
| − | + | ||
| − | + | ||
| − | + | ||
| − | + | ||
| − | + | ||
| − | + | ||
</article> | </article> | ||
| Line 2,636: | Line 1,753: | ||
<p class="note-title">None</p> | <p class="note-title">None</p> | ||
</article> | </article> | ||
| + | |||
<article id="October-17" class="note-item"> | <article id="October-17" class="note-item"> | ||
<h2>October 17</h2><hr> | <h2>October 17</h2><hr> | ||
<p class="note-title">None</p> | <p class="note-title">None</p> | ||
</article> | </article> | ||
| + | |||
<article id="October-18" class="note-item"> | <article id="October-18" class="note-item"> | ||
<h2>October 18</h2><hr> | <h2>October 18</h2><hr> | ||
<p class="note-title">None</p> | <p class="note-title">None</p> | ||
</article> | </article> | ||
| + | |||
<article id="October-19" class="note-item"> | <article id="October-19" class="note-item"> | ||
<h2>October 19</h2><hr> | <h2>October 19</h2><hr> | ||
Revision as of 16:09, 16 October 2018
Wet Lab
July 1
Cloning
- Received and resuspended of the IDT basic part
- Sample
- Enterocin B
- Enterocin 96
- Bovicin HJ50
- Duracin TW49M
- Lacticin Z
- Leucocyclicin Q
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
None
July 5
None
July 6
None
July 7
None
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
None
July 11
None
July 12
None
July 13
None
July 14
None
July 15
Expression
- repared 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
- Duracin 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
- Duracin 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
- Duracin TW49M + pTXB1
Cloning gene of Curcumin biosensor
- Digesting pet30a backbone and ligating it with αS1-casein, GS linker and T7 promoter.
- sample
- pet30a backbone
- gene segment of αS1-casein
- GS linker and T7 promoter
August 14
Cloning
- Cultivation
- Miniprep (Purify Plasmid)
- Sample
- Enterocin B + pTXB1
- Enterocin 96 + pTXB1
- Duracin 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
- Duracin 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
Expression
- Transforming the ligation product of the Curcumin biosensor into E. coli BL21 DE3 Cultivating the culture and inducing it with IPTG.
- Sample
- fresh colony of E. coli BL21 DE3 carrying the backbone pet30a containing αS1-casei and GS linker gene from an overnight plate
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
Chip Production
- Processing the chips with Mua , EtOH , and EDC+NHS mixture, then adding αS1-casein on chips.
- Sample
- αS1-casein protein
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
- Duracin 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
Biosensor Measurement 1- Electrochemical Impedance Spectroscopy
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
- Duracin 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
- Duracin TW49M + pET30a
- Lacticin Z + pET30a
- Leucocyclicin Q + pET30a
August 22
Cloning
- Sequencing plasmid
- Sample
- Enterocin B + pET30a
- Enterocin 96 + pET30a
- Bovicin HJ50 + pET30a
- Duracin TW49M + pET30a
- Lacticin Z + pET30a
- Leucocyclicin Q + pET30a
Biosensor Measurement 2- Differential Pulse Voltammetry
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
- 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
- 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
Cultivation
IPTG Induction
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
- 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
- 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
Cultivation
IPTG Induction
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
