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[seanspotatobusiness]

How stable are the A-overhangs of PCR products? The pGEM-T easy protocol requires that PCR product are purified first, but I'd expect that to eliminate a lot of the T-overhangs. I was made to believe they were pretty delicate. Could that just be a fear instilled by Invitrogen and their pTOPO vector (because I think you don't need to purify the PCR product for that - I don't see any mention of it in their protcol, but it's been over a year since I did it).

Edit: Should be A-overhangs!

Edited by seanspotatobusiness, 14 December 2010 - 06:00 AM.

[bob1]

The A-overhangs do get removed quite quickly, especially if you have some proof-reading polymerase present in the mix.  Typically people use A-overhanged PCR within 24 hours.  The extraction should not remove A-overhangs.