Sticky Problem to a Blunt end - Cloning (Aug/13/2002 )
I have an insert which has got at one end a blunt end restriction site & on the other hand a sticky end restriction site. After PCR amplification of the insert, I am not able to get Proper Fall Out after restriction digestion & therefore no colonies after subsequent cloning & Ligation. Please Help
Where are the restriction sites inthe PCR-product?
If they are in the primers, have you added enough bases at both sides of the restriction sites?