840:153g:Projects/project4/2009/04/07

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Tuesday 4/7

Josh and Casy

  • The PCR products were combined, and digested with the restriction enzymes XbaI and PstI, which were determined to digest the aprE gene so that it will be ligated after the promoter in the plasmid.
  • The digested gene was ran on a gel for one hour in order to separate the two fragments, as discussed previously. However, only a single band could be distinguished. It is possible that the second band seen in the previous gel was due to some other factor not attributable to the DNA.
  • The brightest part of the band was excised and DNA was extracted from the gel using the protocol from QIAquick Gel Extraction Kit. The DNA samples were stored in the freezer, and will be used for ligation.
  • The remaining portion of the band was also excised and stored in the freezer for possible future use.

Derek and Katy

  • Ran a gel electrophorisis on our PCR amplification of wintergreen with three temperatures, two concentrations, and two primers.
  • From the results we found the best combination for PCR amplification of wintergreen.
  • Next class we will do another PCR amplification of wintergreen using primer 8, 1.0 concentration, at 48 degrees since that was the most successful combination.