840:153g:Projects/project4/2009/03/11: Difference between revisions

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* Cells from four areas of the lawns were streaked onto a sectioned LB plate, and incubated.
* 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.
* 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 that number of dimers that can be formed. The mutations, shown in bold, will not interfere with the amino acid sequence.
* 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:
     Forward Primer:
         5’ ATGAATTCGCGGCCGCTTCTAG ATGAG'''G'''AGCAAAAAATTGTGG
         5’ ATGAATTCGCGGCCGCTTCTAG ATGAG(G)AGCAAAAAATTGTGG


     Reverse Primer:
     Reverse Primer:
         5’ ATCTGCAGCGGCCGCTACTAGTA TTATTGTGCAGC'''A'''GCTTGTAC
         5’ ATCTGCAGCGGCCGCTACTAGTA TTATTGTGCAGC(A)GCTTGTAC




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Revision as of 23:15, 11 March 2009

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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’ ATGAATTCGCGGCCGCTTCTAG ATGAG(G)AGCAAAAAATTGTGG

   Reverse Primer:
        5’ ATCTGCAGCGGCCGCTACTAGTA TTATTGTGCAGC(A)GCTTGTAC


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