I did a real time PCR using the Rotor gene Q machine and I analyzed the data using the software. I did't get proper peeks and did't get the Ct values when analysed.
I guess you mean CT value, right? If the CT value is >30-35 your sample does not express the gene of interest or your PCR is not properly working; or your input is too low.
Are you doing genomic qPCR or RT-qPCR? Which species and samples are you trying to analyze? Did you run a conventional PCR and run an agarose gel to check whether your primers are working? How much RNA did you use in your RT or much genomic DNA are you using per reaction? And so on and so forth....?
To answer your question you need to provide more information!
Dear Sabine Strehl, Yes sorry for the mistake. I performed a one step real time PCR with RNA and the RNA was extracted from human adipose tissue.This was my first run and I didn't check if the primers were working. I got degraded RNA but performed the real time PCR anyway. Can it be because of the quality of the RNA?
Yes, it may also be a problem of RNA quality. Test your primers on a tissue, which for sure expresses your gene of interest and use good quality RNA for that; use 2µg of RNA for your RT.
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