Gel Purification -- Anyone else have this problem?
r-mehta at nimr.mrc.ac.uk
Mon May 19 07:31:05 EST 1997
By some eerie co-incidence, we are having the same problems as you are
describing. Briefly, we are trying to clone a 3 kb insert into a 11 kb
vector. To make mattter more interesting, we have to fill in both the
insert and vector DNA's and then perform blunt end ligation. We have
been using Quiagens QuiaexII kit for gel purification. Like you we have
tried every combination without any joy. What we can't understand is
that the colonies we get back are all junk - restriction digests to not
correspond to any of the possible outcomes. We are still scratching our
heads, correction, banging our heads on this one. One of the
possibilities currently persued is using Agarace for gel purification.
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