Elution of DNA from agarose gels
steffen at mbir.bcm.tmc.edu
Wed Nov 13 11:41:47 EST 1991
rick at GENEMAN.WUSTL.EDU (Rick Wilson) writes:
>K.R. Raj writes:
>>I am quite tired of electroeluting DNA out of agarose. Could
>>anyone suggest a better way other than using the geneclean kit ?
>I've had a lot of success with simple phenol extraction of melted
>low-gelling temperature agarose.
I just want to second this, and to add that for many procedures, we
find that we do not need to remove the agarose at all. All of our
ligations, sequencing, and random oligo primed labelling are now done
in low melting point agarose directly. Melt the agarose at 65 degrees
C for five minutes, cool to 37 degrees, mix components, and run the
reaction at the usual temperature. (It doesn't matter if the agarose
hardens again durring the reaction). We have also done restriction
digests this way, but for technical reasons it is usually easier to
purify the reprecipitate the DNA first in this case.
BTW, people in my lab are quite superstitious about sources and lots
of agarose. The only LMP agarose they will use is Genetic Technology
Grade SeaPlaque GTG agarose from FMC BioProducts. I have no
connection with FMC except as a satisfied customer, and I certainly
don't mean to suggest that other brands don't work. I just want to
suggest that if you have difficulties, you might try a different
source of agarose.
Department of Cell Biology, Baylor College of Medicine, Houston TX 77030
Telephone = (713) 798-6655, FAX = (713) 790-0545
Internet = steffen at mbir.bcm.tmc.edu
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