fuzzy pcr - Reply

Varyc varyc.mmcri at OFFICE.MMC.ORG
Fri Dec 2 07:51:54 EST 1994

John Nelson writes:
	I am doing microsatellite amplification from genomic dna and I find
for one of the microsatellites I'm amplifying the bands (around 100-200 bp)
always seem to be fuzzy on a 3% nusieve/1% BRL gel.  I don't think it is the gel
as the marker looks ok and other microsatelites amplified of approx this size
amplify up as more distinct bands.
	Any ideas out there on this?  
John         nelsonj at pbs.dfo.ca

You're mayseeing a phenomenon called stuttering if this microsatellite is of the dinucleotide type.
Stutter is an error made by the polymerase giving products that are of the allele product length +/-
(1,2..). This phenomenon is easily confirmed on a denaturing sequencing gel.
Hope this helps.

Cal Vary

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