purification of 100 bp fragments

Elizabeth Bates bates at cismsun.univ-lyon.fr
Thu Sep 28 10:14:52 EST 1995


jwlafoll at unity.ncsu.edu (James Warren Lafollette) wrote:
>I have been trying to find an efficient method to purify small fragments,
>between 50 - 120 bp, from agarose gels and have not been succesful.  Kits
>like Geneclean only give minimal recovery.  Does anyone have a protocol
>that gives a good, 50-70%, recovery of small fragments.  Thanks, Jamie
>
>

Dear Jamie,
Old fashioned methods like electroelution (am I the only person who still does this?) work wonders with small fragments, you just have to be careful when precipitating. I generally add glycogen when I have fragments less than 500bp or less than 500ng. A tip which works well but adds a more complex step is to electroelute from a TBE-PAGE gel (yes its possible:-)) and you can treat the gel exactly as if it were agarose. I havent really quantified my results but in any case I get more than 50% yield even on a bad day.
Liz.




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