quantifying cDNA by spectrophotometry - (Jun/24/2008 )

hi, biomolecular mates

I'm a novice in quantifying cDNA by spectrophotometry before running a PCR, I wonder if someone can help me by a simple protocole to determine how much in ng do I have in my sample

I will be very grateful if you can help me, I'm opened to any suggestion since I don't have any method

thanks

Unless you have cleaned up your cDNA to remove dNTPs, then you are not able to quantify the cDNA as the dNTPs will interfere with the measurement.

Typically you add a defined quantity of mRNA to the cDNA prep and assume 1:1 conversion of mRNA to cDNA, and that the reaction has gone to completion.