02/24/2011
Kristie Nybo, Ph.D.
After checking my sequence by BLAST, I got 98% identity when sequencing from the forward primer and 93% from the reverse. Is this sufficient to confirm my results?
BioTechniques Molecular Biology Forum brings bench scientists together in an online community to ask for and share advice regarding laboratory problems. This week’s highlighted question comes from the Real-Time qPCR/qRT-PCR Methods forum.
Q: After checking my sequence by BLAST, I got 98% identity when sequencing from the forward primer and 93% from the reverse. Is this sufficient to confirm my results?
A1: First, trim off any bad sequence at the beginning and end of the sequence data and correct any other obvious sequencing artifacts you notice. Then align the forward and reverse sequences and check for… A2: If you have any changes from the known sequence, they may match polymorphisms already in the database. If you find sequence changes such as new polymorphisms, you should…
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