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How to recover small fragment from agaro

John Watson ajwatson at romeo.caltech.edu
Thu Jul 7 14:04:16 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 this kind of purpose (subcloning) it's often easier to do the
subsequent manipulations in the molten gel.  You can restrict and ligate in
the gel, and then transform chemically or clean up the DNA and
electroporate.  If you are intersted I'll send you the protocol I use.

Cheers,

John



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