Gelase instead of Geneclean
rcwieg at ccmail.monsanto.com
Fri Jul 16 08:38:59 EST 1993
In article <1993Jul15.140606.1259 at abo.fi> Mary Metzler,
METZLER at ALA.BTK.UTU.FI writes:
>Has anyone tried Gelase, which is an enzyme which digests LM agarose,
>which eliminates the necessity of using Geneclean? I am tired of the
>low yields and unreliability of Geneclean. I also sometimes have to
>work with big fragments which aren't compatable with Geneclean. Has
>anyone used Gelase? You are supposed to be able to simply ethanol
>precipitate the DNA in your gel slice after digesting with this
>enzyme for an hour. Does anyone know if it works, and if it is
I've had NO problems since I started using Gelase. Excellent recoveries
of all size fragments (up to 16kB tested), great ligations. Its so easy
that I now routinely gel purify cut vectors to drop the background due to
uncut molecules. Yes, the simple EtOH ppt material ligates fine--there is
some contaminating stuff in it, but it doesn't interfere. I've built
about 50 constructs since I started using it and wouldn't change for
anything. Between Gelase and shrimp alkaline phosphatase I went 118/120
on minipreps giving me the right construct last month. Its fast and easy
relative to methods where you have to mess around with spin columns,
freeze-thaw and such. The incubation is an hour, but you have to eat
lunch sometime! 8-)
Yes, it works.
Cheap? A couple bucks per fragment purified (using the "fast", enzyme
intensive protocol)--far less than the cost of repeating a construction.
Seems cheap to me.
Gelase purified DNA labels well by random priming, but I've heard that
people have trouble doing PCR on it. I've never had occasion to try.
No connection to whoever sells it, just a happy user.
rcwieg at ccmail.monsanto.com *****************
Monsanto Corp. Research, Mol. Biol. * Ignotum per *
700 Chesterfield Parkway North * ignotius *
Chesterfield, MO 63198 *****************
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