Mutation detection in DNA fragments

Patrick Lynch
Sun Apr 8 14:35:27 EST 2001


IMHO, SSCP is probably the easiest/fastest way of mutation detection in PCR
fragments.  Briefly, the DNA of interest is amplified (150-300 bp) by PCR,
denatured to single strand DNA by heating in formamide, and electrophoresed
through a high-resolution non-denaturing acrylamide gel.  The presence of a
mutation may result in altered mobilities on the gel.  Using different gel
conditions maximises the chance of finding a mutation.  Detection stratagies
include silver staining, autoradiography (with P32) and fluorescent dye
incorporation.

Orita, M. et al - Genomics. 1989 Nov;5(4):874-9 is one of the first
references I can think of.

Hope this helps,

Patrick


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