840:153g:Projects/project4/2009/02/19

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Thursday 2/19

Josh and Casy

  • The digested plasmid DNA samples (5μL of each) were run on an agarose gel for 30 minutes.
  • The gel indicated that not all of the plasmid was digested, as there were two bands for each sample. This is ready for the aprE ligation, with about 720 ng of digested plasmid available.
  • Some samples may be used to attempt to increase the amount of digested plasmid by repeating the digestion and purification steps, or to isolate the digested plasmid by extracting it from an agarose gel.
  • The samples containing both digested and undigested plasmids can be used in the next step of the experiment.

Derek, Oggie, and Katy

  • Genomic DNA was extracted from both subspecies of B. subtilis following a new protocol which unlike the last protocol utilized a phenol and alcohol purification.
  • There was a problem with the gel, but it appears that DNA was obtained. Our 100 BP ladder showed up, as did our var. Niger sample. However, due to the old gel, our two Globiformous extractions did not run but did show a very bright illumination inside the wells.
  • This will serve as the template DNA for the PCR. We plan to run a gradient to check for best possible anneling temperatures, and then mass produce the aprE gene to inset into our construct.
  • ** We may opt to run a gel with our newly extracted DNA before running our gradient, to be determined.


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