I'm performing some qPCR reactions. My experiment is only about absolute quantification of genomic DNA from some samples. My no template control sample is showing flurorescence at cycle 33. The melt curve shows the same curve of the template samples. Should i assume contamination or there is another possibility like nonspecific products for example? Dimers and hairpin must not be an option according to the primers design. Thanks!
If u are using universal primers in this work, you might easily get DNA contaminant from the water you use to make the reaction mixture. I have experienced that in our lab, even using a freshly bought nanopure water. Perform a simple PCR test using the water as the DNA template before proceeding with your real experiment.
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