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Team:British Columbia/Notebook/6 July 2011
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*Troubleshooting: DMSO could be lowering the Tm of the primers; Repeat the SDM PCR without adding DMSO for one sample, without adding MgCl<sub>2</sub> for another sample. Use the ramping cycling condition previously indicated. | *Troubleshooting: DMSO could be lowering the Tm of the primers; Repeat the SDM PCR without adding DMSO for one sample, without adding MgCl<sub>2</sub> for another sample. Use the ramping cycling condition previously indicated. | ||
**Gel verification of the SDM product shows no bands. Addition of DMSO and MgCl<sub>2</sub> is not the problem. | **Gel verification of the SDM product shows no bands. Addition of DMSO and MgCl<sub>2</sub> is not the problem. | ||
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| + | <b>1,8 Cineole</b> | ||
| + | *Ran another SDM on pg-tps-cin, this time with more PFU. The gel suggests that it worked! Jacob then transformed E. coli with this product. | ||
Revision as of 20:13, 30 July 2011
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July 06 2011
beta-pinene
- Gel verification of the SDM product shows bands of the correct size suggesting that the SDM was successful.
(-)-limonene
- Gel verification of the SDM product shows some lanes with smears, and another with 2 bands suggesting unspecified annealing of primers. SDM failed.
- Troubleshooting: DMSO could be lowering the Tm of the primers; Repeat the SDM PCR without adding DMSO for one sample, without adding MgCl2 for another sample. Use the ramping cycling condition previously indicated.
- Gel verification of the SDM product shows no bands. Addition of DMSO and MgCl2 is not the problem.
1,8 Cineole
- Ran another SDM on pg-tps-cin, this time with more PFU. The gel suggests that it worked! Jacob then transformed E. coli with this product.
