Detection of punctual mutations

Scot Federman scottf19 at nfi.com
Wed Nov 29 23:06:50 EST 1995


In article <Pine.3.89.9511201406.A702-0100000 at argos.uniandes.edu.co>,
Carolina Garcia Imhof <caro-gar at uniandes.edu.co> wrote:

> I am wandering if someone could help me to find alternative methods to 
> find a punctual mutation in a specific DNA sequence. I know that with 
> DGGE I can detect punctual mutations, based on differences of migration 
> in the denaturing gel, using G-C clamps. The problem is that those 
> differences are between mutations of G instead of A or T, but I do not 
> know how to detect a difference if the mutation is of an A instead of T, 
> or a G instead of C, or viceversa, because the rate of denaturation would 
> not change, and there would not be differnces in migration. 
 

You could try SSCP, I have used this technique to screen multiple PCR
products successfully.  There is a lot of literature on this technique,
and theoretically it can distinguish single base mutations regardless of
their type.  It will not necessarily detect 100% of the mutations, but
depending on certain factors (the length of product being used for one) it
isn't bad.

    Scot



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