microsatellites run on the ABI 377

Windows Manager WINMAN at leeds.ac.uk
Tue Oct 28 09:14:19 EST 1997


In article <01bcdf11$f4ac4560$d966c180 at kortej>,
   "John Korte" <kortej at css.orst.edu> wrote:
>Greetings,
>Does anyone out there run microsatellite rxns on an ABI 377?  I am in 
need
>of a protocol, more specifically how much primer you use and how much 
do
>you load on a gel?

I use between 10 and 15 pmoles of fluorescent and non-fluorescent 
primer per 25ul reaction using 25-28 cycles.  I add 1ul of PCR product 
to 4.5ul of loading buffer and load 2ul on the gel.  This normally 
gives 'glowing' bands.

Hope this helps.

J. Moor
Scientific Officer
ICRF Genetic Epidemiology
St James University Hospital 
Leeds
UK



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