I am performing real time pcr for ACTB. I am getting a ct value of 17-20 in my samples. However , i am getting a peak in NTC too with a ct value around 33-35. Also the dissociation curve for NTC shows the peak around same tm as the samples. Is this contamination ?
I believe you are seeing a low level of contamination. Ideally, a NTC should come up as NA, or in the high 30s. This plus your Tm data suggests contamination. I'd systematically go through the components in the NTC wells (water, PCR buffer, ect.) and check for contamination. You could also try using different pipettes.
I believe it is contamination. You probably have to replace all the reagent and change to a different lab to set the PCR up.
sounds like DNA contamination in your primer mix or water or qPCR mix.
I would suggest to replace the nuclease free water and primers and run again. Contamination is very likely in this case. And also try to clean up PCR setup bench.
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