I would like to ask you guys about restriction site.
If I had design a primer for Insert PCR,
With forward primer of AGATCTATGGCGTTGGCGTTG (underline is BglII digestion site)
Notice that I made a mistake and it should be gtAGATCTATGGCGTTGGCGTTG and providing more space for the BglII to hang on it. I wonder whether the enzyme will still do its digestion as the amplicon might not have enough space to hang onto the DNA.
Please give me guidance and advices about this.
Edited by leyhian83, 28 March 2009 - 03:16 PM.