I've heard and read from different sources/ persons that after converting rna into cdna, it is unnecessary to measure its conc. Is it true?
If I moved to rtpcr (qpcr) and take for instance 1ug cdna from each sample, how can I guarantee the accuracy of qpcr results. (as it is possible that each sample has different cdna conc. and it was ignored by ignoring cdna conc. measurement)?
I would quantify your RNA/cDNA at every step if you can. It's important to know just how much you are taking forward to the next step to adjust your protocol and monitor losses.
Bear in mind, however, that some spectrophotometers (Nanodrop/ Epoch) will read left over dNTPs, hexamers, restriction digest material etc and will give you an inaccurate reading.
I'd definitely take the time to run a quick quantification.
I usually didn't, because I was going to normalize to a housekeeping gene anyway.
Try Qbit or Qunti-IT picogreen. There's also the biochips for the agilent bioanalyzer that might work.
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