non-specific amplification in the control tube

Pablo Zaltz pzaltz at yahoo.com
Wed Feb 9 23:24:46 EST 2000


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.






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