R.CARHUAZ at CGIAR.ORG
Fri Nov 27 11:41:53 EST 1998
Using a set of ssr primers I could find some difficult primers in the sense
that they show a nice amplification product on agarose gel, but horrendous
smear on acrylamide gel which makes difficult or imposible to distinguish a
discrete band. With great effort I could succed once and find a very
useful band for my linkage analysis, now I need to apply this difficult
primer to a larger plant-DNA population, however, using the same protocol, I
find a tedious smear rather than the expected band.
Can anyone tell me why this happen?, Does the smear keeps relation with the
amplified genomic region or it is due to the oligo sequence itself?. I
don'I think the smear might due to PCR components or DNA purity as they work
nicely with other primers.
But the MOST important, please tell me how to rid me off this undesirable
(to say the least) smear.
Thanks a lot in advance for any input.
Rina Carhuaz Ambia
International Potato Center
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