840:153g:Projects/project23/2012/10/04

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Cloning of the OmcF gene Main project page
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  • On Tuesday we worked on finishing our DNA extraction form the Geobacter bacteria.
  • We spun our sample down to pour out the extra supernatant, suspended it in 500 microL TE and 2 microL of RNase, this was then incubated
  • After letting our sample incubate we did a process of 4 DNA extractions in hopes to rid our sample of all proteins and other extra materials besides pure DNA. This was done with 2 extractions of phenol and 2 extractions with chloroform. These were spun to separate the layers of phenol/chloroform and DNA. We discarded these after spinning it down.
  • We then added 1mL of ethanol and 40 microL of Na-acetate and let it precipitate until Thursday
  • On Thursday we resuspended our pellet in 50microL of H2O and then began our set up for running a PCR
  • We created an appropriate liquid mixture for our get- using agrose and TBE buffer, then added EtBr and poured into our bed to create gel.
  • We created a digest with our DNA using EcoR1, buffer, and H2O and incubated
  • We used 3 wells:
                  Well #3- undigested DNA
                  Well #4- 100Bp ladder
                  Well #5- digested DNA
  • Results are here and are read from left to right 3,4,5 Image:DNA Gel Oct 04 2012.jpg



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