Team:HKUST/Notebook

From 2010.igem.org

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<p class="content"><strong>14/06/2010</strong><br />
+
 
-
   E.coli DH10B competent cells were successfully transformed with BioBrick<br />
+
  <p class="content">14/06/2010 <br />
-
  BBa_I746101 which contains agrC.</p>
+
   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.  
-
<p class="content"><strong>25/06/2010</strong> <br />
+
  </p>
-
  PMH4 containing mCherry and PBI121 conatining gusA reporter gene were<br />
+
 
-
  successfully transferred into E.coli DH10B competent cells.</p>
+
  <p class="content">25/06/2010  
-
<p class="content"><strong>12/07/2010</strong> <br />
+
  <br />PMH4 containing mCherry and PBI121 conatining gusA reporter gene were successfully transferred into E.coli DH10B competent cells.
-
  Fusion PCR of agrC-mCherry was performed and products were confirmed by agarose<br />
+
  </p>
-
  gel electrophoresis.</p>
+
 
-
<p class="content"><strong>30/07/2010</strong> <br />
+
  <p class="content">12/07/2010  
-
  Genomic DNA of Lactobacillus. Plantarum WCFS1 was extracted. Next, PCR of<br />
+
  <br />Fusion PCR of agrC-mCherry was performed and products were confirmed by agarose gel electrophoresis.
-
  plnB from the genomic DNA was conducted and products were confirmed by<br />
+
  </p>
-
   agarose gel electrophoresis.</p>
+
 
-
<p class="content"><strong>10/08/2010</strong><br />  
+
  <p class="content">30/07/2010  
-
   Midi-prep of shuttle vector pMG36e was performed and extracted plasmids were<br />
+
  <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.
-
  confirmed by enzyme digestion test.</p>
+
  </p>
-
<p class="content"><strong>16/08/2010</strong> <br />
+
    
-
  PCR of gusA from PBI121 was conducted and products were confirmed by agarose<br />
+
  <p class="content">21/07/2010
-
  gel electrophoresis.</p>
+
  <br />Successfully extract  plasmid pMG36e out from filter paper.
-
<p class="content"><strong>17/08/2010</strong> <br />
+
  </p>
-
  Fusion PCR of agrC-plnB was performed and products were confirmed by agarose gel<br />
+
 
-
   electrophoresis.</p>
+
  <p class="content">22/07/2010
-
<p class="content"><strong>20/08/2010</strong> <br />
+
  <br /><em>E.coli</em> DH10B competent cells were successfully transformed with pMG36e
-
  Double-strand DNA of plnA promoter (124bp) was obtained. By following the<br />
+
  </p>
-
  protocol of fusion PCR, annealing of the forward primer (79bp) with the reverse<br />
+
 
-
  primer (73bp) and the extension were accomplished.</p>
+
  <p class="content">28/07/2010  
-
<p class="content"><strong>26/08/2010</strong> <br />
+
  <br />Enzyme digested the vector  and confirmed plasmid pMG36e.
-
  agrC-plnB was ligated into pBluescript KS (+). Ligation products were transferred<br />
+
  </p>
-
  into E.coli DH10B competent cells by heat shock transformation. Ligation was<br />
+
    
-
  confirmed by colony PCR and plasmid digestion test.</p>
+
  <p class="content">10/08/2010
-
<p class="content"><strong>13/09/2010</strong> <br />
+
  <br />Midi-prep of shuttle vector pMG36e was performed and extracted plasmids were confirmed by enzyme digestion test.</p>
-
  agrC was ligated into pBluescript KS (+). Ligation products were transferred<br />
+
  <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>
-
  into E.coli DH10B competent cells by heat shock transformation. Ligation was<br />
+
  <p class="content">17/08/2010 <br />Fusion PCR of agrC-plnB was performed and products were confirmed by agarose gel electrophoresis.</p>
-
  confirmed by colony PCR and plasmid digestion test.</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"><strong>24/09/2010</strong> <br />
+
  <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>
-
   gusA was ligated into backbone pSB1C3. Ligation products were transferred<br />
+
  <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>
-
  into E.coli DH10B competent cells by heat shock transformation. Ligation was<br />
+
  <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>
-
  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">12/10/2010<br />GUS assay with the substrate  X-Gluc. E.coli containing PBI121 gave more gusA expression than control group.
 +
  </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>
</div>
</div>

Revision as of 14:14, 15 October 2010

Team: HKUST

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.

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.

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.

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.

28/08/2010
Primer (*****-*****) arrived and repare primers for ligation. Double strand DNA of FLAG-tag, both part 1&2 of signal peptide and DD13-RIP was obtained.

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.

13/09/2010
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.

26/09/2010
Sequencing of the insert was obtained by DNA sequencing to further confirm the ligation product.

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.

12/10/2010
GUS assay with the substrate X-Gluc. E.coli containing PBI121 gave more gusA expression than control group.

13/10/2010
GUS assay with the substrate 4-NPG. E.coli containing PBI121 gave more gusA expression than control group.

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