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ligating blunt ends of a linearized plasmid, is kinase necessary? - (Mar/04/2013 )

A quick cloning question:

I want to delete part of a plasmid. I double digest with to REs which result in sticky ends. I gel-purify the desired linearized fragment. I blunt-end with T4 polymerase and the ligate the ends together by T4 ligase.

Before the ligation, do I need to use T4 kinase to add phosphate groups to 5' ends?

thank you



No, T4 ligase is sufficient for blunt end ligation. The provided ATP will provide the phosphate linkage.