| | Re: what are they in my EMSA?
Thanks very much!
the double-strand NF-KB consensus sequence(5'agt tga ggg gac ttt ccc agg c3‘) has 22bp in length. the concentration of my nuclear extrat was about 2ug/ul,so 2ul volum of nuclear extract was added to the binding action which included 10xbinding buffer(2ul) poly(1ul) 50%glycerol(1ul) 40%NP-40(1ul) Mgcl2(1ul) EDTA(1ul) and had total volum of 20ul. i'm sorry i don't know the copoments of the cell lysates of PIERCE kit.
the pellet was difficult to be dispersed after the nucli lysate(50ul) was added to the nuclear pellet .Do you have similar details and how to solve it?