Difference between revisions of "Team:Rock Ridge Virginia/Results"

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<p> We picked 5 colonies from each plate and we purified the plasmids using mini-preps. We checked the plasmid purity using agarose gels, all of the colonies had the plamid inside. We did not sent the plasmids for sequencing due to the cost, we just selected one of the five colonies and did a PCR using Gibson Assembly primers. We had a lot of difficulty doing the PCRs because the melting temperature between the primers was not similar. It took us quite some time to get all of amplicons ready, especially WSP to which we had to add b-mercaptoethol to get a good yield.  
 
<p> We picked 5 colonies from each plate and we purified the plasmids using mini-preps. We checked the plasmid purity using agarose gels, all of the colonies had the plamid inside. We did not sent the plasmids for sequencing due to the cost, we just selected one of the five colonies and did a PCR using Gibson Assembly primers. We had a lot of difficulty doing the PCRs because the melting temperature between the primers was not similar. It took us quite some time to get all of amplicons ready, especially WSP to which we had to add b-mercaptoethol to get a good yield.  
  
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<li> Our first attempt at PCR
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<p><img src="https://static.igem.org/mediawiki/2015/9/9c/Rock_Ridge_PCR_first_attempt.jpeg"></li><h6>
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<li> Optimized PCR amplicons :)
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<p><img src="https://static.igem.org/mediawiki/2015/4/49/Rock_Ridge_PCR_gel.jpeg"></li><h6>
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Revision as of 00:50, 19 September 2015

Project Results

Parts

Once we received our parts from IDT we transformed the plasmids in DH5-alpha cells in ampicillin resistant plates.All of our plates (OspA, WSP and GFP) had many colonies due to the fact that we added 3 times the amount of DNA since we had some problems with transformation in our practice rounds. The positive plates were positive and the negative plates were negative.

We picked 5 colonies from each plate and we purified the plasmids using mini-preps. We checked the plasmid purity using agarose gels, all of the colonies had the plamid inside. We did not sent the plasmids for sequencing due to the cost, we just selected one of the five colonies and did a PCR using Gibson Assembly primers. We had a lot of difficulty doing the PCRs because the melting temperature between the primers was not similar. It took us quite some time to get all of amplicons ready, especially WSP to which we had to add b-mercaptoethol to get a good yield.

  • Our first attempt at PCR

  • Optimized PCR amplicons :)

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