Agarose to determine heterozygotes

Terry Delaney tpd4 at cornell.edu
Wed Nov 29 18:17:57 EST 1995


In article <e-anderson-2211951413240001 at e-mac-56.ski.mskcc.org>,
e-anderson at ski.mskcc.org (Eric C. Anderson) wrote:

> In article <Pine.A32.3.91.951122111948.48216A-100000 at umabnet.ab.umd.edu>,
> "Mary P. Remington" <mremingt at umabnet.ab.umd.edu> wrote:
> 
> > Is there an agarose type gel on the market that will separate ~20 base 
> > pair differences in a PCR product?  Any help or avenues to explore will 
> > be appreciated.  Thanks, Mary
> 
> FMC makes an agarose called MetaPhor(tm) which they claim is almost as
> good as PAGE for separating closely spaced products.  i've never looked
> into the price but i'm sure that it's a little expensive.
> 
> eric


Metaphor agarose works very well to resolve small differences in size. 
For DNA 100-500 bp, 2% is reliably achieved, meaning your 20 bp
polymorphism should be easily visualized (if total <1 kbp).  A 1:1 mixture
(or higher) of FMC NuSieve:ordinary high quality agarose (eg FMC GTG) will
resolve a 20 bp difference if the total size is <500 bp (tl.
agarose=1.5%/TBE).  Metaphor is somewhat expensive*, which more apparent
when making the 2-4% gels needed.  This is offset somewhat by reusing the
gels, which I have done up to four times.  

*'95 catalog lists 100g/$203 for Metaphor.

Terry Delaney
Cornell


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  Terrence P. Delaney           Department of Plant Pathology
  Cornell University                           (607) 255-7856
  334 Plant Science Building               FAX (607) 255-4471
  Ithaca, NY 14853-4203                      tpd4 at cornell.edu
   http://ppathw3.cals.cornell.edu/ppath/Delaney/Delaney.html
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