ligation in LMA

Clemens Suter-Crazzolara un691cs at genius.embnet.dkfz-heidelberg.de
Wed Aug 17 05:47:33 EST 1994


Hi,

up to now, I have ligated in low-melt agarose quite succesfully,
but I must admit with low efficiency. I realized that the protocol
that I am using is at least five years old: I mix the fragments,
add water and ligation buffer and 2 (!) units of T4 ligase. All in
60 microliters (!), the total volume of agarose being below 10 micro-
liters.
Surely this method can be improved ! Any useful hints out there ?
(I don't want to purify the fragments from agarose: too much hassle).

clemens suter-crazzolara, uni Heidelberg




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