Registry/Measurement kit/Notebook/2007-7-31: Difference between revisions
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(New page: ==E0240== *Noticed that during the scarring PCR, a point substitution was made. *Decided to try a site-directed mutagenesis on the biobricked product. *Ordered two pairs of primers: **...) |
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Revision as of 14:19, 31 July 2007
E0240
- Noticed that during the scarring PCR, a point substitution was made.
- Decided to try a site-directed mutagenesis on the biobricked product.
- Ordered two pairs of primers:
- First pair - mutation in the ends of the primers:
- CGAAAGATCCCAACGAAAAGAGAGACCACATGGTCCTTCTTGAGTTTGTAACAGC, E0240_mut_E_F (78.8)
- CTCTCTTTTCGTTGGGATCTTTCGAAAGGGCAGATTGTGTGGACAGG, E0240_mut_E_R (79.7)
- Second pair - mutation in the middle of the primers:
- CCTTTCGAAAGATCCCAACGAAAAGAGAGACCACATGGTCCTTCTTGAGTTTG, E0240_mut_M_F (80)
- ACCATGTGGTCTCTCTTTTCGTTGGGATCTTTCGAAAGGGCAGATTG, E0240_mut_M_R (79.3)
- First pair - mutation in the ends of the primers:
I2057
- Re-transformed both versions with ccdB into DB3.1
Misc
- Made chemically competent DB3.1 cells.
