molecular cloning - (May/10/2009 )
I am planning to do cloning of a GST fusion protein in PGEX vector. I need to do restriction digestion of PCR amplified product with BamH1 and EcoR1 (restriction site present at 5'ends of primers ). I want to know how many additional bases BamH1 needed in addition to restriction site at the end of primer for efficient digestion or just restriction site is enough without additional bases.
You do need to add additional bp around the restriction site. Please check NEB technical guide on their website.
vikrant saa on May 10 2009, 04:44 AM said:
addition of extra bases is always advisable. as a general rule, I add extra 3 bases.