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Team:Stockholm/19 August 2010
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Received the sequencing results for [[media:M-sod1_premix_19aug.txt|m-SOD 1]], [[media:M-sod2_premix_19aug.txt|m-SOD 2]], [[media:M-yCCS1_premix_19aug.txt|m-yCCS 1]] and [[media:M-yCCS4_premix_19aug.txt|m-yCCS 4]] from Eurofins. Ran a nucleotide BLAST (BLASTn) alignment to verify successful site-directed mutagenesis. | Received the sequencing results for [[media:M-sod1_premix_19aug.txt|m-SOD 1]], [[media:M-sod2_premix_19aug.txt|m-SOD 2]], [[media:M-yCCS1_premix_19aug.txt|m-yCCS 1]] and [[media:M-yCCS4_premix_19aug.txt|m-yCCS 4]] from Eurofins. Ran a nucleotide BLAST (BLASTn) alignment to verify successful site-directed mutagenesis. | ||
| - | *'''SOD 1:''' Failed | + | *'''SOD 1:''' Failed ([[media:BLASTn_m-SOD1_19aug.txt|BLASTn results]]) |
**Mutation still present | **Mutation still present | ||
| - | *'''SOD 2:''' Success | + | *'''SOD 2:''' Success ([[media:BLASTn_m-SOD2_19aug.txt|BLASTn results]]) |
| - | **Mutation removed | + | **Mutation removed |
| - | *'''yCCS 1:''' Success | + | *'''yCCS 1:''' Success ([[media:BLASTn_m-yCCS1_19aug.txt|BLASTn results]]) |
**EcoRI site removed | **EcoRI site removed | ||
| - | *'''yCCS 4:''' Success | + | *'''yCCS 4:''' Success ([[media:BLASTn_m-yCCS4_19aug.txt|BLASTn results]]) |
**EcoRI site removed | **EcoRI site removed | ||
SOD 2 sample and glycerol stock will be saved, while SOD 1 will be discarded.<br /> | 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'''. | 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 10:10, 19 August 2010
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Andreas
Site-directed mutagenesis of SOD & yCCS
Sequencing results
Received the sequencing results for m-SOD 1, m-SOD 2, m-yCCS 1 and m-yCCS 4 from Eurofins. Ran a nucleotide BLAST (BLASTn) alignment to verify successful site-directed mutagenesis.
- SOD 1: Failed (BLASTn results)
- Mutation still present
- SOD 2: Success (BLASTn results)
- Mutation removed
- yCCS 1: Success (BLASTn results)
- EcoRI site removed
- yCCS 4: Success (BLASTn results)
- 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.
