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Team:British Columbia/Notebook/Week 6
From 2011.igem.org
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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.
