Results and Discussion LR2

Results and Discussion LR2 - Results and Discussion...

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Results and Discussion Polymerase Chain Reaction is a process used to amplify copies of certain DNA sequences. We use this process to amplify the luxAB gene from the Vibrio fischeri DNA. We used the Vibrio fischeri DNA obtained from earlier in this lab to use in PCR. After running PCR, we run our sample through a gel electrophoreses cycle to see if the PCR reaction worked. In the gel we should see two bands, one in the V. fischeri DNA well and one in the 500/501 positive control well. When comparing the intensity of these bands to the digested HindIII control, which was also run in the gel, we are able to determine the amount of PCR product we have produced. We estimate that we have about 500ng of DNA in the V. fischeri band which we divide by 5ul, then multiply the result by 45ul. Using this calculation we conclude that we have approximately 450ng of PCR product present. Now that we know the amount of PCR product present we can continue onto the purification process. The PCR products we obtained are not purified and need to be cleaned from salts, primers, and enzymes that may still be present. We use the Qiagen MinElute PCR Purification kit to perform the purification process. We assume that we recovered 100% of the DNA run through the MinElute PCR Purification process. While we were isolating the luxAB gene from V. fischeri DNA using the methods mentioned earlier, we were also
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This note was uploaded on 06/10/2009 for the course BICD 655698 taught by Professor Gustafson-brown,cindy during the Winter '09 term at UCSD.

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Results and Discussion LR2 - Results and Discussion...

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