Hello everybody,
I am running a PCR to amplify a 200 bp fragment of the Cauliflower
Mosaic Virus (CaMV) 35S promoter. I get a non-specific amplification
between 100 and 500 bp in the negative control tube (just mastermix +
water).
I have descarted any kind of contamination. Could be possible that in
the situation of primers self-amplification? Do you have any suggestion
to explain the problem? Can you suggest a web site where to analize this
parameters?
My PCR protocol inculude two well-known 35S promoter primers:
35S-1 gctcctacaaatgccatca
35S-2 agtgggattgtgcgtca
Run: 94 C/3min
94 C/20 sec ]
56 C/30 sec ] 35 cycles
72 C/60 sec ]
72 C/3 min
Thank in advance,
Pablo.