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How to recover small fragment from agaro

gc genecutl at mendel.berkeley.edu
Thu Jul 7 17:59:10 EST 1994


In article <199407071416.XAA29594 at inetnif.niftyserve.or.jp>,
HGF02633 at NIFTYSERVE.OR.JP (=?ISO-2022-JP?B?GyRAQFBETT1TPCMbKEo=?=) wrote:

> Dear Netters
> I separated the 100 bp, 50 bp, and 30 bp DNA fragments with NuSieveGTG
> low melting point agarose gel. I want to subclone the 100 bp 
> fragment. I tried to purify the 100 bp fragment. However, the recovery 
> is very poor (less than 30 %). 
> After electrophoresis, I excised the gel band of 100 bp DNA,  melt the
> gel at 65 C,  did phenol extraction, and precipitated the DNA with 
> ethanol (in 0.1 M NaCl). Does anybody know how to recover small DNA 
> from agarose gel efficiently. Please teach me.
> Sincerely yours,
> ================================================================
> Dr.Toshiharu Ishizuka
> Department of Biochemistry, Chiba University, School of Medicine

It sounds like the main problem is the ethanol precipitation.  The amounts
of DNA your playing with are going to be fairly small and if you want
to have efficient precipitation you should add a carrier like glycogen
or RNA.




-- 
--gc

I know you like rock 'n' roll. Have you considered overdosing
on heroin, like many of your pop-star favorites?  --Life in Hell (M.G.)



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