Check Clones for Correct Mutation

  1. Send for sequencing, preferably with primers reading both forward and reverse of the mutation
  2. Go to http://www.ncbi.nlm.nih.gov/blast/bl2seq/wblast2.cgi then click on tblastx on the tabs
  3. Paste dna sequence in top box
  4. Paste sequence, gi number or accession number for the refseq protein sequence into the lower box
  5. Click BLAST
  6. Look at the sequences and check for expected mutation
  7. Check both forward and reverse sequences