trouble with pcr,urgent

turkish_scientist_f · #1 (**permalink**) 09-17-2007, 11:04 AM

Hello all

i have a big problem,i have smearing problem with the pcr..there is a massive smearing that i cant get bands at all..i tried everything ,changed annealing temp., cycles,mgcl2 concentration... and tried hot start taq. but couldnt get bands..i must solute it ,cause 3000 dna samples waiting for that test

is there anyone 2 help me?

Ammie · #2 (**permalink**) 09-17-2007, 12:33 PM

Have you tried to dilute your cDNA? (to maybe 50 ng/ul) To much template have given smears for me.

cintiadp · #3 (**permalink**) 10-07-2007, 02:30 AM

Did you check the quality of your RNA or DNA?

turkish_scientist_f · #4 (**permalink**) 11-09-2007, 08:10 AM

its not about my dna contentration or quality,i can work the pcr perfectly with another primers,but just one couple of primers are smearing,and i have no another choice of that analise

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trouble with pcr,urgent

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