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How to recover small fragment from agaro

FJ van der Wal vdwal at bio.vu.nl
Fri Jul 8 03:04:01 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.
For recovery of a 64 bp (PCR) fragment from a 2% agarose gel I used the
protocol in TIG V7 (4) p 109 , 1991.'Fast recovery of DNA from agarose gels
by centrifugation through blotting paper'. Subsequently I precipitated the
DNA with lineair polyacrylamide, which is very efficient: NAR (1990) V18(2)
p 378. I used it also for larger fragments, and it always worked for me.
Hope this helps.
-- 
Fimme Jan van der Wal
dep. Molecular Microbiology
Vrije Universiteit Amsterdam
de Boelelaan 1087
1081 HV Amsterdam, NL
vdwal at bio.vu.nl



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