Team:Stockholm/19 August 2010

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Revision as of 09:58, 19 August 2010 by AndreasConstantinou (Talk | contribs)


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
    • 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.