small sized restricted bands couldnt be visualized - restriction enzymes problems (Mar/04/2005 )

hello

thanks alot JIM
i performed a testing PCR yeaterday but my annealing is 51
tomorrow i well incubate with Mbo 11 for 2 hours and test on agarose 45 to see what well appear
thanks for ur help
i well tell u all what happens

Lula,
How did your MTHFR run go? Anealing at 51 C might be ok; I've seen protocols that call for 55C. I Run Factor V Leiden, Prothrombin II, and MTHFR on the same thermocycler program and 65 fits them all pretty well.
Jim

hello

im thinking about raising the annealing temperature from 51 to 57 because i have been getting a smear in the PCR BAND lately
(my primers Tm is 64 )
as regard the restriction ,,,still no luck
but i didnt give up yet