recovery of small fragment
llintott at ACS.UCALGARY.CA
Thu Jul 7 10:20:41 EST 1994
> 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
> 1-8-1, Inohana, Chuo-ku, Chiba, 260 JAPAN
> FAX +81-43-226-2041
> QFH03407 at niftyserve.or.jp
> tishizuk at twics.com
Try using Bio101's mermaid kit. I have used it to purify a
fragment as small as 166 bp from polyacrylamide gel. I know
someone else who purified a 100 bp fragment for cloning with the
same kit. It worked great in both cases.
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