RNA to cDNA conversion - question relating to reverse transcription rates from RNA to cDNA (May/15/2008 )

Hi

I was wondering if anyone can help me out with a question.

If a set amount of RNA was added to an RT reaction (e.g. 2ug in a 20ul volume) would I be able to estimate or calculate the concentration of final amount of cDNA that is synthesised?

I am very aware that cDNA can be quantified using kits like picogreen etc...
but I want to know if there is a generally accepted mathematical approach....

please keep your answer as simple as possible!

thanks in advance.

Madu

Since RT reaction is not cyclical it would give you the same number of cDNA as the mRNA you input.
1 million mRNA = 1 million cDNA

consider these too:

1. Not same number as total RNA molecule, mRNA only.
2. If you use OligodT for priming, theoretically, you will get the same number of cDNA molecules as are intact mRNA molecules.
3. If you use random primers, the number of cDNA molecules will be much larger.
4. Amount (not number) of cDNA may very, generally lower than mRNA.

hth/