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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==I2057== | ==I2057== | ||
*Re-transformed both versions with ccdB into DB3.1 | *Re-transformed both versions with ccdB into DB3.1 | ||
==I2055 and I2056== | |||
*Letting biobrick PCR run overnight. | |||
==T9002== | |||
*Re-tried, transformed ligation of F2620. | |||
==Misc== | ==Misc== | ||
*Made chemically competent DB3.1 cells. | *Made chemically competent DB3.1 cells. | ||
Latest revision as of 14:57, 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
I2055 and I2056
- Letting biobrick PCR run overnight.
T9002
- Re-tried, transformed ligation of F2620.
Misc
- Made chemically competent DB3.1 cells.
