CASTING METAPHOR AGAROSE GELS

GERALDINE KAMINSKI gzk at ollamh.ucd.ie
Thu Jun 13 11:40:54 EST 1996


In article <31BD9671.4D02 at acs.tamu.edu>, pjb6578 at ACS.TAMU.EDU ("Perry J. Bushong") says:
>
>I have had a difficult time trying to get consistent 
>results running microsatellites on 4% Metaphor 
>agarose gel.  The gels always run crooked.  One side 
>of my 34 lane, 3mm gel always runs faster than the 
>other.  Yet there is no pattern to it; sometimes the 
>left side runs faster, sometimes the right, and 
>sometimes the middle.  I thought the problem was due 
>to an unlevel casting tray, but I always use a 
>leveler to make sure.  I then thought maybe my 
>agarose was hardening before it spread out so I tried 
>submerging the Bio-rad casting tray under warm water 
>(about 65degC) the same temp as the agarose being 
>poured. I even tried using a 3% gel but got the same 
>results. I have never had so much trouble casting a 
>gel before. 
>Does anyone out know any tricks to casting Metaphor 
>gels consistently?  Thanks.
>Perry

We use Metaphor regularly and have never had a problem 
with it.  We found it was difficult to get it to dissolve 
by heating in the microwave it is best to melt on a heating
block or by decreasing the power on the microwave to 6 instead
of 10.  It does congeal very quickly and I must admit to pouring
it almost immediately without cooling it to 65 (we don't measure
the temperature).  Our casting trays have sides on them and only 
have to be taped at the ends, this gives us straight bands.
Also are you remembering to keep them at 4 deg for half an hour 
before running them?
Hope this is of help 
Geraldine



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