just received a message from a user with this question:
I have been working on RAPD (Randomly Amplified Polymormphic DNA).
Im working under the DBT project as finding the molecular markers for growth in Fish.
I was getting good results with Peqlab Primus 98 thermocycler machine with the following concentrations:
DNA 15ng= 2 micro litre
dNTP 5mM=1 micro litre
taq bufferA10x = 2 micro litre(which comes along with the Taq polymerase from the company GENIE)
Taq polymerase (GENIE)3u/microlitre= 0.4micro litre
MgCl2 (25mM)= 1.2 micro litre primers screened and using (SIGMA COMPANY)2pM=12micro litre
OPG 9 OPP16 OPB17 OPA7 OPF9 OPG4 OPF 10
But now suddenly i have started getting smear of DNA.
Ive tried with different concentrations of DNA, MgCl2, Taq. But still I'm not getting results. Can you please help me out in solving this problem.