Team:Stockholm/19 August 2010
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+ | ===Site-directed mutagenesis of SOD & yCCS=== | ||
+ | ====Sequencing results==== | ||
+ | |||
+ | Received the sequencing results from Eurofins. Ran a nucleotide BLAST (BLASTn) alignment to verify successful site-directed mutagenesis. | ||
+ | *'''SOD 1:''' Failed | ||
+ | **Mutation still present | ||
+ | *'''SOD 2:''' Success | ||
+ | **Mutation removed | ||
+ | *'''yCCS 1:''' Success | ||
+ | **EcoRI site removed | ||
+ | *'''yCCS 4:''' Success | ||
+ | **EcoRI site removed | ||
+ | |||
+ | SOD 2 sample and glycerol stock will be saved, while SOD 1 will be discarded.<br /> | ||
+ | Unfortunately, BLAST results also discovered 4 (!) identical mutations in both yCCS sequences. A protein BLAST (BLASTp) alignment of the translated yCCS sequences revealed all four mutations as '''silent mutations'''. |
Revision as of 09:47, 19 August 2010
Andreas
Site-directed mutagenesis of SOD & yCCS
Sequencing results
Received the sequencing results from Eurofins. Ran a nucleotide BLAST (BLASTn) alignment to verify successful site-directed mutagenesis.
- SOD 1: Failed
- Mutation still present
- SOD 2: Success
- Mutation removed
- yCCS 1: Success
- EcoRI site removed
- yCCS 4: Success
- EcoRI site removed
SOD 2 sample and glycerol stock will be saved, while SOD 1 will be discarded.
Unfortunately, BLAST results also discovered 4 (!) identical mutations in both yCCS sequences. A protein BLAST (BLASTp) alignment of the translated yCCS sequences revealed all four mutations as silent mutations.