840:153g:Projects/project4/2009/03/26: Difference between revisions

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     Forward Primer:
     Forward Primer:
         5’ TAGAATTCGCGGCCGCTTCTAG ATGAG(G)AGCAAAAAATTGTGG
         5’ TAGAATTCGCGGCCGCTTCTAG ATGAG(G)AGCAAAAAATTGTGG
     Reverse Primer:
     Reverse Primer:
         5’ ATCTGCAGCGGCCGCTACTAGTA TTATTGTGCAGC(A)GCTTGTAC
         5’ ATCTGCAGCGGCCGCTACTAGTA TTATTGTGCAGC(A)GCTTGTAC

Revision as of 13:31, 26 March 2009

MoldBusters' Journal <html><img src="/images/9/94/Report.png" border="0" /></html> Main project page
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Thursday 3/26

Oggie

  • Diluted primers from 100μM to 10μM concentrations.
  • Setup a gradient for the 3 different forward primers in order to obtain optimal annealing temperature.
  • Each primer was tested individually as well as in combination with the reverse primer to test for dimers.

Josh and Casy

  • Discussed PCR running conditions for Tuesday when new primers arrive.
  • The PCR will be run with 1μL and 0.1μL of DNA. The temperatures that will range from 45°C, 50°C, 55°C, and 60°C.
  • The new primers contain a point mutation that will eliminate hairpin formation.
    Forward Primer:
       5’ TAGAATTCGCGGCCGCTTCTAG ATGAG(G)AGCAAAAAATTGTGG

    Reverse Primer:
       5’ ATCTGCAGCGGCCGCTACTAGTA TTATTGTGCAGC(A)GCTTGTAC


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