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Team:British Columbia/Notebook/Week 6
From 2011.igem.org
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==1,8-cineole== | ==1,8-cineole== | ||
| - | [[File:UbcTubesonice.jpg| | + | [[File:UbcTubesonice.jpg| thumb | right | 200px]] |
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. | 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. | ||
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==Beta-pinene== | ==Beta-pinene== | ||
| - | [[File:ubcthreeinlab.jpg| thumb | left | | + | [[File:ubcthreeinlab.jpg| thumb | left | 300px]] |
All 3 transformation plates (P2SDM1, P2SDM2, P2SDM3) have colonies! Marianne set up overnight cultures, mini-prepped them and sent them for sequencing. Hopefully the SDM worked and the beta-pinene synthase gene no longer has illegal cut sites. | All 3 transformation plates (P2SDM1, P2SDM2, P2SDM3) have colonies! Marianne set up overnight cultures, mini-prepped them and sent them for sequencing. Hopefully the SDM worked and the beta-pinene synthase gene no longer has illegal cut sites. | ||
Revision as of 05:59, 8 August 2011
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Contents |
Week 6: July 10-16
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.
Beta-pinene
All 3 transformation plates (P2SDM1, P2SDM2, P2SDM3) have colonies! Marianne set up overnight cultures, mini-prepped them and sent them for sequencing. Hopefully the SDM worked and the beta-pinene synthase gene no longer has illegal cut sites.
(-)-limonene
SDMs have failed thus far. More 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.
Again, gel verification shows no bands. More troubleshooting: use a different cycling condition.
Again, no bands. Something is amiss... Vicki might start pulling out her hairs.
IDI1, HMG2 metabolic genes
Yeast colonies were found for HMG2, p330 and Cin, but not IDI1. The yeast transformed with IDI1 did not grow on a URA- plate. This may be because the nutrient marker was mutated in the plasmid, but this would not affect PCR to amplify the gene out. As a precaution, IDI1 was re-amplified.
