Hi,
We have a small test run for a set of RNA samples with 3 fluorophores, FAM, HEX, and Cy5. FAM and HEX are for test samples while Cy5 is with internal control. Surprisingly, for some of our samples, there are no Cq values but have an amplification curve in the RFU plot. Another sample doesn't have any RFU plot but has a Cq value.
Is this possible?
Alok Tiwary what brand of PCR machine are you using? This could be a software issue. Can you include some pictures of your plots where you have this issue?
Hello
You may set the baseline to both automatic or manual. Sometimes you need to change the default of your software, especially when you have diverse type of samples or even this is your first run. Anyway you may see the real results when you do an agarose gel.
This is purely due to the baseline correction error. You adjust it manually by looking at your qPCR curves, just above the noise level. You would see the Ct values for all the samples showing amplification. Also check that all samples showing amplification also shows corresponding melting curve.
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