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?
ligating blunt ends of a linearized plasmid, is kinase necessary?
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