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Template:Team:Edinburgh/Notebook/Week05
From 2012.igem.org
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<div class="entry-text"> | <div class="entry-text"> | ||
<p> | <p> | ||
| - | < | + | <b> ccm transformation</b><br/> |
| + | competent E coli were transformed with ccm plasmids | ||
| + | |||
</p> | </p> | ||
</div><!-- /article-text --> | </div><!-- /article-text --> | ||
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<div class="entry-text"> | <div class="entry-text"> | ||
<p> | <p> | ||
| - | < | + | <b> ccm transformants subculture</b><br/> |
| + | ccm transformed cells that lost RFP were subcultured. | ||
</p> | </p> | ||
</div><!-- /article-text --> | </div><!-- /article-text --> | ||
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<div class="entry-text"> | <div class="entry-text"> | ||
<p> | <p> | ||
| - | < | + | <b> liquid cultures preparation</b><br/> |
| + | subcultures of ccm transformed cells were used to inoculate 3ml LB; samples were left to grow overnight<br/><br/> | ||
| + | <b> primers preparation </b><br/> | ||
| + | Stock solution (500 pmol/ul, solution in EB) and working solution (10 pmol/ul 1 in 50 silution of stock solution in water) of primers were prepared for nfsl, cymA, mtrA and mtrCAB gene primers. <br/><br/> | ||
| + | <b> Kod PCR of Shewanella </b><br/> | ||
| + | Kod Pcr was performed for potential Shewanella oneidensis strain using 3 sets of primers: cymA, mtrA and mtrCAB. Gel analysis indicated no results. | ||
</p> | </p> | ||
</div><!-- /article-text --> | </div><!-- /article-text --> | ||
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<div class="entry-text"> | <div class="entry-text"> | ||
<p> | <p> | ||
| - | < | + | <b> Herculase PCR for Shewanella </b><br/> |
| + | Herculase PCR was performed for potential Shewanella oneidensis strain using 3 sets of primers: cymA, mtrA and mtrCAB. Gel analysis indicated no results. | ||
</p> | </p> | ||
</div><!-- /article-text --> | </div><!-- /article-text --> | ||
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<div class="entry-text"> | <div class="entry-text"> | ||
<p> | <p> | ||
| - | < | + | <b> Shewanella PCR </b> |
| + | Multiple PCRs were prepared:</br> | ||
| + | Kod PCR for 16S ribosomal DNA for potential Shewanella strain (strains was later identified as different bacteria)<br/> | ||
| + | Taq PCR for 3 potential Shewanella strains using mtrA primers (one of the strains showed positive result at expected 1 kb)<br/><br/> | ||
| + | <b> Half-cell setup</b> | ||
| + | Half-cell was autoclaved, filled with liquid LB medium and inocluated with potential Shewanella strain. The Half-cell was sealed and left for incubation over the weekend. The initial voltage readout was 0.4 mV. | ||
</p> | </p> | ||
</div><!-- /article-text --> | </div><!-- /article-text --> | ||
Revision as of 10:41, 1 August 2012
ccm transformants subculture
ccm transformed cells that lost RFP were subcultured.
liquid cultures preparation
subcultures of ccm transformed cells were used to inoculate 3ml LB; samples were left to grow overnight
primers preparation
Stock solution (500 pmol/ul, solution in EB) and working solution (10 pmol/ul 1 in 50 silution of stock solution in water) of primers were prepared for nfsl, cymA, mtrA and mtrCAB gene primers.
Kod PCR of Shewanella
Kod Pcr was performed for potential Shewanella oneidensis strain using 3 sets of primers: cymA, mtrA and mtrCAB. Gel analysis indicated no results.
Herculase PCR for Shewanella
Herculase PCR was performed for potential Shewanella oneidensis strain using 3 sets of primers: cymA, mtrA and mtrCAB. Gel analysis indicated no results.
Shewanella PCR
Multiple PCRs were prepared:
Kod PCR for 16S ribosomal DNA for potential Shewanella strain (strains was later identified as different bacteria)
Taq PCR for 3 potential Shewanella strains using mtrA primers (one of the strains showed positive result at expected 1 kb)
Half-cell setup
Half-cell was autoclaved, filled with liquid LB medium and inocluated with potential Shewanella strain. The Half-cell was sealed and left for incubation over the weekend. The initial voltage readout was 0.4 mV.
