qieex problems

j.r.stephen mbi071 at abdn.ac.uk
Wed Feb 28 10:43:03 EST 1996


I have also seen the same problem, and though it costs a little, the following
has been foolproof so far. Simply dilute the DNA eluted from the resin in 0.45
mls TE, and pass through a Microcon 100 column. When fully spun through, discard
the TE, and pass 0.45 mls of the highest quality water you have through the
column. Elute in as little as 5 microlitres or whatever suits. This ligates
beautifully. Though I haven't tried it, any kind of ultrafiltration should do
the trick of getting rid of whatever the contaminant is. Using a Millipor type
VS membrane as described in Maniatis would probably work, though you may have
problems keeping the volume down.

I came to this method because precipitation/ washing did not seem to help.

Good luck,

John Stephen



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