840:153g:Projects/project4/2009/04/09

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Thursday 4/9

Josh and Casy

  • Ran a gel of the extracted aprE gene from Tuesday in order to confirm purity and determine concentration. The second elution samples contained little to no DNA. The first elution samples, however, contained a very high concentration of DNA, about 13 ng/μL.
  • Using this concentration of gene and the contration of the digested plasmid, the ligation was setup in ratios of 1:1, 2:1, and 1:5 (Based on molar ratios of plasmid to gene).
  • The ligation contained the appropriate amount of plasmid and gene, as well as ligase and buffer. Controls to test self-ligation will be done Friday.
  • 800 mL of LB+Amp plates were prepared.

Oggie and Katy

  • Setup a PCR using old DNA strains from B. subtilis globiformis and niger in combination with our newly synthesizes aprE primers, and used original aprE primers with the DNA from B. subtilis strain 168
  • Results will hopefully reveal whether our initial failures were due to poorly synthesized primers, different DNA between the strains, or possibly both
  • Also setup a PCR to mass produce the wintergreene gene that Katy and Derek are working on


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