Team:British Columbia/Notebook/Week 6

From 2011.igem.org

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==July 11 2011==
 +
<b>beta-pinene</b>
 +
*Transformation plates taken out.
 +
*All 3 plates (P2SDM1, P2SDM2, P2SDM3) have colonies.
 +
*Set up overnight culture for miniprepping.
 +
 +
<b>(-)-limonene</b>
 +
*SDM PCR has failed thus far.
 +
*Troubleshooting: Use less pfu as the glycerol content in the enzyme could alter the reaction - prepared 3 samples, each with 0.5uL pfu, 1ul pfu, and 2ul pfu, respectively. Also prepare a sample using 0.5 taq polymerase only. Also increased the annealing temperature and elongation time.
 +
 +
==July 12 2011==
 +
<b>beta-pinene</b>
 +
*Miniprep one colony from each of the three plates.
 +
 +
<b>(-)-limonene</b>
 +
*Gel verification shows no bands
 +
*Troubleshooting: Use a different cycling condition
 +
 +
 +
<b>1,8-cineole</b>
 +
*After 5 SDMs, transformations, and minipreps, Jacob should now have ECORI-less, SPEI-less, XBAI-less, PSTI-less pg-tps-cin and pgxe-tps-cin. The gel of the restriction digest has the correct number of bands. Sequencing is the next step to confirm this result.
 +
 +
==July 13 2011==
 +
<b>beta-pinene</b>
 +
*sent miniprepped samples for sequencing.
 +
 +
<b>(-)-limonene</b>
 +
*Gel verification of SDM products shows no bands.

Revision as of 15:54, 3 August 2011

Team: British Columbia - 2011.igem.org

UbcTubesonice.jpg

July 11 2011

beta-pinene

  • Transformation plates taken out.
  • All 3 plates (P2SDM1, P2SDM2, P2SDM3) have colonies.
  • Set up overnight culture for miniprepping.

(-)-limonene

  • SDM PCR has failed thus far.
  • Troubleshooting: Use less pfu as the glycerol content in the enzyme could alter the reaction - prepared 3 samples, each with 0.5uL pfu, 1ul pfu, and 2ul pfu, respectively. Also prepare a sample using 0.5 taq polymerase only. Also increased the annealing temperature and elongation time.

July 12 2011

beta-pinene

  • Miniprep one colony from each of the three plates.

(-)-limonene

  • Gel verification shows no bands
  • Troubleshooting: Use a different cycling condition


1,8-cineole

  • After 5 SDMs, transformations, and minipreps, Jacob should now have ECORI-less, SPEI-less, XBAI-less, PSTI-less pg-tps-cin and pgxe-tps-cin. The gel of the restriction digest has the correct number of bands. Sequencing is the next step to confirm this result.

July 13 2011

beta-pinene

  • sent miniprepped samples for sequencing.

(-)-limonene

  • Gel verification of SDM products shows no bands.
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