Gavin Fleming 108GFLEM at WITSVMA.WITS.AC.ZA
Fri May 20 02:19:47 EST 1994

Resolution of microsatellites with Metaphor agarose

I am attempting to resolve dinucleotide repeat microsatellites on Metaphor
agarose.  The idea is that it will be simpler and less hazardous than
polyacrylamide and hot labelling.

I get fragments from 70 - 110 bp at the loci I am investigating, but no matter

what I have tried, I cannot get tight enough bands to resolve 2bp differences
between alleles.   I have used up to 10V/cm, 4,5% agarose, narrow-tooth comb,
running in a cold-room, to no avail.

Can anyone suggest an improvement or two, or should I give up on the claim
that Metaphor can resolve down to 1-2% differences in fragment size?

In the meantime, I'm getting ready to switch to polyacrylamide and silver

Gavin Fleming
Genetics Dept.
Wits University

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