microsatellites

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
staining.

Gavin Fleming
Genetics Dept.
Wits University
Johannesburg



More information about the Methods mailing list