isolating 197bp fragment

Dr. Duncan Clark Duncan at nospam.demon.co.uk
Thu Nov 23 04:39:57 EST 2000


In article <Pine.LNX.4.21.0011222207070.31533-100000 at biochem.iisc.ernet.
in>, the eminent SURESH KUMAR K.G. at BIOSCI/MRC Human Genome Mapping
Project Resource Centre wrote
>You can use the glass wool elution method.
>I think a protocol for this has been posted earlier in the group.
>Any way here it goes..
>
>1.Pierce a small 0.5ml eppendorf with a needle (18g) and plug it with
>  silianized glass wool. 
>
>2.Keep it in a large eppendorf and spin at 6500 rpm for 5min 
>
>3.Cut the gel(1.0%)slice into very small pieces.
>
>4.Put it back into the eppendorf with glass wool and place it in a fresh
>  large tube.

I would avoid the use of glass wool and instead use aquarium filter
wool. It is autoclavable and has been reported to work better
(Biotechniques (?) a long time ago). If you have high salt around, your
DNA can bind to the glass wool. Not surprising given the no. of DNA
purification kits using silica/glass fibre discs to purify DNA.

Duncan
-- 
The problem with being on the cutting edge is that you occasionally get 
sliced from time to time....

Duncan Clark
GeneSys Ltd.
Tel: +44(0)1252376288
FAX: +44(0)8701640382
http://www.dnamp.com
http://www.genesys.demon.co.uk






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