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extraa bases before RE site - Reply urgent!! (May/31/2011 )

Hello everyone!

I have a basic doubt what if I dont keep any extra bases before the restriction site.. Would this create any problems in cloning of PCR products..

Please reply fast! As I am facing this problem.. but not sure whether it is because of this..

-AmrutaK-

If you are digesting your pcr product before ligation, maybe. Some enzymes need few extra bases to 'land on'.
If you are cloning pcr ptoduct into simple vector as pGem t easy, no.
Also depends which polymerase you are using, some eat the ends of primers if you take your time seting up the reaction.

-kajmak-