We want to do qPCR from single cells extracted from patch-clamp recordings.
As the RNA content of each cell will be very small and to prevent further degradation, i will not quantify it, instead synthesizing cDNA out of everything I collected as soon as collected. Then I'll do a pre-amplification step and then qPCR.
My question is how to best quantify the cDNA for qPCR input?
Should I use qubit with a ssDNA kit?
Or can I use nanodrop and assume the primer and dntp concentrations are equal among samples?
Should I make a standard curve out of my control gene in the qPCR to quantify the samples?
Should I quantify it before or after the pre-amplification step?
I was thinking of going for both the nanodrop and standard curve after the pre-amplification (I dont have easy access to a qubit) and comparing before choosing
hope you can help me find the best way
Thank you!
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