840:153g:Projects/project4/2009/03/11

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Wednesday 3/11

Josh, Katy, and Oggie

  • The Bacillus subtilis showed significant growth, forming a lawn of cells around both locations the filter paper was placed.
  • Cells from four areas of the lawns were streaked onto a sectioned LB plate, and incubated.
  • Two liquid cultures containing TSB broth were inoculated with the growing cells.
  • The primers that have been used were determined to form hairpins and dimers with highly negative ΔG values. This was found to be primarily due to the sequences of the prefix and suffix. However, the following primers are proposed, which eliminate hairpins at operating temperature, and reduces the number of dimers that can be formed. The mutations, shown in parantheses, will not interfere with the amino acid sequence.
   Forward Primer:
        5’ TAGAATTCGCGGCCGCTTCTAG ATGAG(G)AGCAAAAAATTGTGG
   Reverse Primer:
        5’ ATCTGCAGCGGCCGCTACTAGTA TTATTGTGCAGC(A)GCTTGTAC