Check Clones for Correct Mutation
From Bridges Lab Protocols
- Send for sequencing, preferably with primers reading both forward and reverse of the mutation
- Go to http://www.ncbi.nlm.nih.gov/blast/bl2seq/wblast2.cgi then click on tblastx on the tabs
- Paste dna sequence in top box
- Paste sequence, gi number or accession number for the refseq protein sequence into the lower box
- Click BLAST
- Look at the sequences and check for expected mutation
- Check both forward and reverse sequences