I have 2 concentrations from different samples, Nanodrop for each are 715.5 ng/ul and 467.5 ng/ul and approximately 2 ul was used for each sample.
I am very confused on how to get and equal concentration/amount from both in order to make cDNA to then use for RT-PCR.
I've looked up the protocol we are using (MMLV RT) but don't see anything on how to actually do the calculations.
I'm really really new and everything I keep reading about converting RNA into equal amounts for cDNA is very confusing and I don't know if there is a general equation available.
Please help. I don't know what to do...
Hi,
By using a simple calculation you can maintain equal amount of RNA to be converted to cDNA. Just divide the amount of template you require by the ng/ul of RNA. Suppose you require 1000ng templare for each sample, so divide 1000 by each sample's RNA concentration.
For example, in this case 1000/715.5= 1.39ul of RNA and for second one 1000/467.5=2.14ul of RNA template will give you 1000 ng template forboth samples. This is how you can ensure equal template amount in all your samples.
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