Difference between revisions of "Team:Dalhousie Halifax NS/Experiments"
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| − | <h4> Preparation of Cellulose Media </h4> | + | <h4><a href="https://static.igem.org/mediawiki/2016/1/19/T--Dalhousie_Halifax_NS--CelluloseMediaProtocol.pdf"> Preparation of Cellulose Media </a></h4> |
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<p>Congo Red Cellulose media can be used to isolate bacteria that degrade cellulose. It does this because the only carbon source in the media is cellulose. It uses Whatman Paper and gelatin.</p> | <p>Congo Red Cellulose media can be used to isolate bacteria that degrade cellulose. It does this because the only carbon source in the media is cellulose. It uses Whatman Paper and gelatin.</p> | ||
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| − | <h4> 16S Colony PCR </h4> | + | <h4><a href="https://static.igem.org/mediawiki/2016/c/c5/T--Dalhousie_Halifax_NS--ColonyPCRProtocol.pdf"> 16S Colony PCR </a></h4> |
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<p>Colony PCR has many applications including; verifying that an insert has been successfully incorporated in plasmid constructs, insert orientation, and species determination of unknown samples. The later was the reason for our use of Colony PCR. After streaking fecal sample onto cellulose plates, we picked colonies of different morphologies to find out which species or genus they were. We did this by using primers that targeted conserved sequence found in bacteria so we could amplify the DNA of almost all bacteria. </p> | <p>Colony PCR has many applications including; verifying that an insert has been successfully incorporated in plasmid constructs, insert orientation, and species determination of unknown samples. The later was the reason for our use of Colony PCR. After streaking fecal sample onto cellulose plates, we picked colonies of different morphologies to find out which species or genus they were. We did this by using primers that targeted conserved sequence found in bacteria so we could amplify the DNA of almost all bacteria. </p> | ||
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| − | <h4> ExoSapIt and DNA Cleanup </h4> | + | <h4><a href="https://static.igem.org/mediawiki/2016/6/69/T--Dalhousie_Halifax_NS--ExoSapItProtocol.pdf"> ExoSapIt and DNA Cleanup </a></h4> |
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<p>Instead of PCR clean up, we used ExoSap It. ExoSap It allows for an enzymatic clean-up of left over dNTPs and single stranded DNA. It is a very quick procedure as the only activation ExoSap It needs is a 15 minute 37°C incubation and a 15 minute 80 °C inactivation.</p> | <p>Instead of PCR clean up, we used ExoSap It. ExoSap It allows for an enzymatic clean-up of left over dNTPs and single stranded DNA. It is a very quick procedure as the only activation ExoSap It needs is a 15 minute 37°C incubation and a 15 minute 80 °C inactivation.</p> | ||
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Revision as of 12:12, 10 October 2016
Experiments
High Through-put Sequencing
Isolation of Bacteria
Congo Red Cellulose media can be used to isolate bacteria that degrade cellulose. It does this because the only carbon source in the media is cellulose. It uses Whatman Paper and gelatin.
Colony PCR has many applications including; verifying that an insert has been successfully incorporated in plasmid constructs, insert orientation, and species determination of unknown samples. The later was the reason for our use of Colony PCR. After streaking fecal sample onto cellulose plates, we picked colonies of different morphologies to find out which species or genus they were. We did this by using primers that targeted conserved sequence found in bacteria so we could amplify the DNA of almost all bacteria.
Instead of PCR clean up, we used ExoSap It. ExoSap It allows for an enzymatic clean-up of left over dNTPs and single stranded DNA. It is a very quick procedure as the only activation ExoSap It needs is a 15 minute 37°C incubation and a 15 minute 80 °C inactivation.
