Team:HKUST/Notebook

From 2010.igem.org

(Difference between revisions)
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<div class="ex">
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<div id="leftpart"> <h1>Lab Notebook</h1></div>
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<div id="leftpart">
 +
<div id="date">14/06/2010 </div>
 +
<div id="logbook">BioBrick BBa_I746101 was extracted from 96  wells plates. <em>E.coli</em> DH10B competent  cells were successfully transformed with BioBrick BBa_I746101 which contains  agrC. </div>
 +
</div>
-
  <p class="content">14/06/2010 <br />
+
 
-
  BioBrick BBa_I746101 was extracted from 96 wells plates. <em>E.coli</em> DH10B competent cells were successfully transformed with BioBrick BBa_I746101 which contains  agrC.  
+
<div id="leftpart">
-
  </p>
+
<div id="date">25/06/2010 </div>
 +
<div id="logbook">PMH4 containing mCherry and  PBI121 conatining gusA reporter gene were successfully transferred into <em>E.coli</em>  DH10B competent cells. </div>
 +
</div>
 +
 
 +
<div id="leftpart">
 +
<div id="date">12/07/2010 </div>
 +
<div id="logbook">Fusion PCR of agrC-mCherry  was performed and products were confirmed by agarose gel electrophoresis.</div>
 +
</div>
 +
 
 +
 
 +
<div id="leftpart">
 +
<div id="date">21/07/2010 </div>
 +
<div id="logbook">Successfully extract plasmid pMG36e out from filter paper.</div>
 +
</div>
 +
 
 +
<div id="leftpart">
 +
<div id="date">22/07/2010 </div>
 +
<div id="logbook"><em>E.coli</em> DH10B competent cells were successfully transformed with pMG36e.</div>
 +
</div>
    
    
-
  <p class="content">25/06/2010
+
<div id="leftpart">
-
  <br />PMH4 containing mCherry and  PBI121 conatining gusA reporter gene were successfully transferred into E.coli  DH10B competent cells.
+
<div id="date">28/07/2010 </div>
-
  </p>
+
<div id="logbook">Enzyme digested the vector  and confirmed plasmid pMG36e.</div>
-
 
+
</div>
-
  <p class="content">12/07/2010  
+
   
-
  <br />Fusion PCR of agrC-mCherry  was performed and products were confirmed by agarose gel electrophoresis.
+
<div id="leftpart1">
-
  </p>
+
<div id="date">30/07/2010 </div>
-
 
+
<div id="logbook">Genomic DNA of <em>Lactobacillus. Plantarum </em>WCFS1was extracted. Next, PCR of plnB from the genomic DNA was conducted and products were confirmed by agarose gel  electrophoresis.</div>
-
  <p class="content">30/07/2010
+
</div>
-
  <br />Genomic DNA of <em>Lactobacillus. Plantarum </em>WCFS1was extracted. Next, PCR of plnB from  the genomic DNA was conducted and products were confirmed by agarose gel  electrophoresis.
+
-
  </p>
+
-
 
+
-
  <p class="content">21/07/2010
+
-
  <br />Successfully extract  plasmid pMG36e out from filter paper.
+
-
  </p>
+
-
 
+
-
  <p class="content">22/07/2010
+
-
  <br /><em>E.coli</em> DH10B competent cells were successfully transformed with pMG36e
+
-
  </p>
+
-
 
+
-
  <p class="content">28/07/2010
+
-
  <br />Enzyme digested the vector  and confirmed plasmid pMG36e.
+
-
  </p>
+
-
 
+
-
  <p class="content">10/08/2010
+
-
  <br />Midi-prep of shuttle vector  pMG36e was performed and extracted plasmids were confirmed by enzyme digestion  test.</p>
+
-
  <p class="content">16/08/2010 <br />PCR of <em>gusA</em> from PBI121 was conducted and products were confirmed by agarose gel electrophoresis.</p>
+
-
  <p class="content">17/08/2010 <br />Fusion PCR of agrC-plnB was  performed and products were confirmed by agarose gel electrophoresis.</p>
+
-
  <p class="content">28/08/2010 <br />Primer (*****-*****)  arrived and repare primers for ligation. Double strand DNA of FLAG-tag, both  part 1&amp;2 of signal peptide and DD13-RIP was obtained.</p>
+
-
  <p class="content">20/08/2010 <br />Double-strand DNA of plnA  promoter (124bp) was obtained. By following the protocol of fusion PCR, annealing  of the forward primer (79bp) with the reverse primer (73bp) and the extension  were accomplished.</p>
+
-
  <p class="content">26/08/2010 <br />agrC-plnB was ligated into  pBluescript KS (+). Ligation products were transferred into <em>E.coli </em>DH10B competent cells by heat shock  transformation. Ligation was confirmed by colony PCR and plasmid digestion  test.</p>
+
-
  <p class="content">13/09/2010 <br />agrC was ligated into  pBluescript KS (+). Ligation products were transferred into <em>E.coli</em> DH10B competent cells by heat shock  transformation. Ligation was confirmed by colony PCR and plasmid digestion  test. </p>
+
-
  <p class="content">20/09/2010 <br />Performed three-way  ligation of SP with plasmid pMG36e.</p>
+
-
  <p class="content">22/09/2010 <br />Colony PCR shown that the insert was successfully ligated into plasmid pMG36e.</p>
+
-
  <p class="content">26/09/2010 <br />Sequencing of the insert was obtained  by DNA sequencing to further confirm the ligation product. </p>
+
-
  <p class="content">24/09/2010 <br />
+
-
  gusA was ligated into  backbone pSB1C3. Ligation products were transferred into <em>E.coli </em>DH10B competent cells by heat shock transformation. Ligation  was confirmed by colony PCR and plasmid digestion test.</p>
+
-
  <p class="content">29/09/2010 <br />
+
-
  Two constructs agrC and agrC-plnB were transferred from pBluescript KS (+) to shuttle vector pMG36e. The ligation products were confirmed by colony PCR and enzyme digestion.</p>
+
-
<p class="content">05/10/2010<br />Sequencing of four constructs: agrC in pBluescript KS (+), agrC-plnB in pBluescript KS (+), agrC in
+
-
pMG36e and agrC-plnB in pMG36e. All of them were finally confirmed.
+
-
  </p>
+
-
<p class="content">11/10/2010<br />mCherry was ligated into pBluescript SK (+). Some colonies on the plate showed red color expressed by mCherry.
+
<div id="leftpart">
-
  </p>
+
<div id="date">10/08/2010 </div>
 +
<div id="logbook">Midi-prep of shuttle vector  pMG36e was performed and extracted plasmids were confirmed by enzyme digestion  test.</div>
 +
</div>
-
  <p class="content">12/10/2010<br />GUS assay with the substrate  X-Gluc. E.coli containing PBI121 gave more gusA expression than control group.<br />agrC-plnB (for localization test) was ligated into pBluescript SK (+). Ligation product was
+
<div id="leftpart">
 +
<div id="date">16/08/2010 </div>
 +
<div id="logbook">PCR of <em>gusA</em> from PBI121 was conducted and products were confirmed by  agarose gel electrophoresis.</div>
 +
</div> 
 +
 
 +
<div id="leftpart">
 +
<div id="date">17/08/2010 </div>
 +
<div id="logbook">Fusion PCR of agrC-plnB was  performed and products were confirmed by agarose gel electrophoresis.</div>
 +
</div> 
 +
 
 +
<div id="leftpart1">
 +
<div id="date">20/08/2010 </div>
 +
<div id="logbook">Double-strand DNA of plnA  promoter (124bp) was obtained. By following the protocol of fusion PCR, annealing  of the forward primer (79bp) with the reverse primer (73bp) and the extension  were accomplished.</div>
 +
</div> 
 +
 
 +
<div id="leftpart1">
 +
<div id="date">26/08/2010 </div>
 +
<div id="logbook">agrC-plnB was ligated into  pBluescript KS (+). Ligation products were transferred into <em>E.coli </em>DH10B competent cells by heat shock  transformation. Ligation was confirmed by colony PCR and plasmid digestion  test.</div>
 +
</div>
 +
 
 +
<div id="leftpart">
 +
<div id="date">28/08/2010 </div>
 +
<div id="logbook">Primers arrived and prepareed primers for ligation. Double strand DNA of FLAG-tag, both  part 1&amp;2 of signal peptide and DD13-RIP was obtained.</div>
 +
</div> 
 +
 
 +
 
 +
<div id="leftpart1">
 +
<div id="date">12/09/2010 </div>
 +
<div id="logbook">agrC was ligated into  pBluescript KS (+). Ligation products were transferred into <em>E.coli</em> DH10B competent cells by heat shock  transformation. Ligation was confirmed by colony PCR and plasmid digestion  test.</div>
 +
</div> 
 +
 +
<div id="leftpart">
 +
<div id="date">20/09/2010 </div>
 +
<div id="logbook">Performed three-way  ligation of SP with plasmid pMG36e.</div>
 +
</div> 
 +
 
 +
<div id="leftpart">
 +
<div id="date">22/09/2010 </div>
 +
<div id="logbook">Colony PCR shown that the  insert was successfully ligated into plasmid pMG36e..</div>
 +
</div> 
 +
 
 +
<div id="leftpart1">
 +
<div id="date">24/09/2010 </div>
 +
<div id="logbook"> gusA was ligated into  backbone pSB1C3. Ligation products were transferred into <em>E.coli </em>DH10B competent cells by heat shock transformation. Ligation  was confirmed by colony PCR and plasmid digestion test.</div>
 +
</div> 
 +
 
 +
<div id="leftpart">
 +
<div id="date">26/09/2010 </div>
 +
<div id="logbook">Sequencing of the insert was obtained  by DNA sequencing to further confirm the ligation product.</div>
 +
</div> 
 +
 
 +
<div id="leftpart1">
 +
<div id="date">29/09/2010 </div>
 +
<div id="logbook"> Two constructs agrC and agrC-plnB were transferred from pBluescript KS (+) to shuttle vector pMG36e. The ligation products were confirmed by colony PCR and enzyme digestion.</div>
 +
</div> 
 +
 
 +
<div id="leftpart">
 +
<div id="date">05/10/2010 </div>
 +
<div id="logbook">Sequencing of four constructs: agrC in pBluescript KS (+), agrC-plnB in pBluescript KS (+), agrC in
 +
pMG36e and agrC-plnB in pMG36e. All of them were finally confirmed.</div>
 +
</div> 
 +
 
 +
<div id="leftpart">
 +
<div id="date">11/10/2010 </div>
 +
<div id="logbook">mCherry was ligated into pBluescript SK (+). Some colonies on the plate showed red color expressed by mCherry.</div>
 +
</div> 
 +
 
 +
<div id="leftpart1">
 +
<div id="date">12/10/2010 </div>
 +
<div id="logbook">GUS assay with the substrate  X-Gluc. E.coli containing PBI121 gave more gusA expression than control group.<br />
 +
agrC-plnB (for localization test) was ligated into pBluescript SK (+). Ligation product was
confirmed by colony PCR and enzyme digestion.
confirmed by colony PCR and enzyme digestion.
-
  </p>
 
-
  <p class="content">13/10/2010<br />GUS assay with the substrate 4-NPG. E.coli containing PBI121 gave more gusA expression than control group.</p>
 
-
<p class="content">15/10/2010<br />Construct agrC-plnB-mCherry was built by ligating mCherry into pBluescript SK (+) containing
 
-
agrC-plnB already.</p>
 
</div>
</div>
 +
</div> 
 +
 +
 +
<div id="leftpart">
 +
<div id="date">13/10/2010 </div>
 +
<div id="logbook">GUS assay with the substrate 4-NPG. E.coli containing PBI121 gave more gusA expression than control group.
 +
</div>
 +
</div>
 +
 +
<div id="leftpart">
 +
<div id="date">15/10/2010 </div>
 +
<div id="logbook">Construct agrC-plnB-mCherry was built by ligating mCherry into pBluescript SK (+) containing
 +
agrC-plnB already.
 +
</div>
 +
</div>
 +
</div>
</div>
</body>
</body>
</html>
</html>

Revision as of 05:37, 23 October 2010

Team: HKUST

Lab Notebook

14/06/2010
BioBrick BBa_I746101 was extracted from 96 wells plates. E.coli DH10B competent cells were successfully transformed with BioBrick BBa_I746101 which contains agrC.
25/06/2010
PMH4 containing mCherry and PBI121 conatining gusA reporter gene were successfully transferred into E.coli DH10B competent cells.
12/07/2010
Fusion PCR of agrC-mCherry was performed and products were confirmed by agarose gel electrophoresis.
21/07/2010
Successfully extract plasmid pMG36e out from filter paper.
22/07/2010
E.coli DH10B competent cells were successfully transformed with pMG36e.
28/07/2010
Enzyme digested the vector and confirmed plasmid pMG36e.
30/07/2010
Genomic DNA of Lactobacillus. Plantarum WCFS1was extracted. Next, PCR of plnB from the genomic DNA was conducted and products were confirmed by agarose gel electrophoresis.
10/08/2010
Midi-prep of shuttle vector pMG36e was performed and extracted plasmids were confirmed by enzyme digestion test.
16/08/2010
PCR of gusA from PBI121 was conducted and products were confirmed by agarose gel electrophoresis.
17/08/2010
Fusion PCR of agrC-plnB was performed and products were confirmed by agarose gel electrophoresis.
20/08/2010
Double-strand DNA of plnA promoter (124bp) was obtained. By following the protocol of fusion PCR, annealing of the forward primer (79bp) with the reverse primer (73bp) and the extension were accomplished.
26/08/2010
agrC-plnB was ligated into pBluescript KS (+). Ligation products were transferred into E.coli DH10B competent cells by heat shock transformation. Ligation was confirmed by colony PCR and plasmid digestion test.
28/08/2010
Primers arrived and prepareed primers for ligation. Double strand DNA of FLAG-tag, both part 1&2 of signal peptide and DD13-RIP was obtained.
12/09/2010
agrC was ligated into pBluescript KS (+). Ligation products were transferred into E.coli DH10B competent cells by heat shock transformation. Ligation was confirmed by colony PCR and plasmid digestion test.
20/09/2010
Performed three-way ligation of SP with plasmid pMG36e.
22/09/2010
Colony PCR shown that the insert was successfully ligated into plasmid pMG36e..
24/09/2010
gusA was ligated into backbone pSB1C3. Ligation products were transferred into E.coli DH10B competent cells by heat shock transformation. Ligation was confirmed by colony PCR and plasmid digestion test.
26/09/2010
Sequencing of the insert was obtained by DNA sequencing to further confirm the ligation product.
29/09/2010
Two constructs agrC and agrC-plnB were transferred from pBluescript KS (+) to shuttle vector pMG36e. The ligation products were confirmed by colony PCR and enzyme digestion.
05/10/2010
Sequencing of four constructs: agrC in pBluescript KS (+), agrC-plnB in pBluescript KS (+), agrC in pMG36e and agrC-plnB in pMG36e. All of them were finally confirmed.
11/10/2010
mCherry was ligated into pBluescript SK (+). Some colonies on the plate showed red color expressed by mCherry.
12/10/2010
GUS assay with the substrate X-Gluc. E.coli containing PBI121 gave more gusA expression than control group.
agrC-plnB (for localization test) was ligated into pBluescript SK (+). Ligation product was confirmed by colony PCR and enzyme digestion.
13/10/2010
GUS assay with the substrate 4-NPG. E.coli containing PBI121 gave more gusA expression than control group.
15/10/2010
Construct agrC-plnB-mCherry was built by ligating mCherry into pBluescript SK (+) containing agrC-plnB already.

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