No PCR product for genotyping - (Apr/10/2006 )

Dear All!!!!!

I've never met such problem before....

I got some hundreds of DNA samples for genotyping...all reactions work before....BUT I don't got any product.

I can see the DNA in the agarose gel..., today I measured the DNA conc. (1,5ug/ul-good!), 260/280 ratio (1,8-1,95-good!), everything is all right, but no band after PCR, and no results after real time PCR.....


(I've changed dNTP mix, primer dliution, water, etc... )


HELP!



Thanks!

Maybe a stupid suggestion but have you tried a new vial of enzyme? I spent the last two weeks trying to amplify a gene and altered everything over and over...it turned out that the enzyme was not good anymore.

Are you using a new set of primers?

Are you sure about the DNA concentration. You cannot overload a PCR reaction with DNA. Maybe recheck your concentration.