Protocol Online logo
Top : Forum Archives: : Molecular Cloning

What EDTA does in annealing of DNA and vector? - (Oct/05/2007 )

I am using Ligation independent cloning (LIC) for insertion of target DNA into vector (pEMT-11).
From the protocol manual, the next step after mixing T4 polymease digested vector and DNA for annealing, is to add 1ul of EDTA and stand for 5 mins.
I 've been asking why do I add it and how important is it?

Or anyone know what function EDTA is (what for)? Or when do you use it?


what i know about EDTA is Antibacterial


Apart from being a useful buffer, EDTA is mostly known as a metal chelator. Depending on the concentration (which you didn't give... wink.gif ), I suppose it could also act to stabilise the non-covalently joined insertion prior to transfecting into the cell.