cDNA quantification by non-PCR method - measuring RT efficiency (Jun/05/2008 )
hello everyone. I'm trying to measure RT efficiency, more specifically the amount of cDNA I get from a given RNA amount. Need to try different conditions and incubation times, so need a way to be able to measure and compare.
Preferably I'd like to use something faster than PCR, ie, measuring cDNA, but as there'll still be RNA in the sample, ssDNA measurment methods will be skewed by the presence of ssRNA in the mix.
So, here's the final question. Has anyone use Quant-iT™ OliGreen ® ssDNA Reagent and Kit to measure cDNA, and does it measure RNA?
any other suggestions will be greatly appreciated.
usually when creating cDNA i use RNase H (included in the kit i use), that should reduce the amount of RNA when you measure your cDNA