"

From 2010.igem.org

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.