CASTING METAPHOR AGAROSE GELS

Hopkins, Charlotte hopkinsc at lincoln.ac.nz
Mon Jun 17 15:09:33 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

Welcome to one of the joys of Metaphor.  We try and avoid it, due to wavy 
bands.  A hint that we have  found does help, if you do need to use it, is to 
have the running buffer chilled as well as the gel.  For some reason we get 
better bands if we put the running buffer in the fridge as well as the gel, 
for at least 1/2 a hour, and sometimes we even run it in there.  We may still 
get some slight waves but at least it is readable. 
Hope this helps

Charlotte Cameron
Lincoln University
Canterbury
New Zealand





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