Checking cDNA concentration - Is it possible to check? (Aug/03/2007 )
I was wondering if there was a way to check cDNA concentration for use in QRT-PCR. Right now I check my RNA concentration and then take 2 ug of RNA to make cDNA.
-proteo-
You can check it using a spectrophotometer. Just clean it up after reverse transcription (using for example PCR purification kit) if it is necessary.
-Tomasz-
You have to amke sure that you don't have any dNTPs or digested RNA left in there, they will confound your measurements... pure dNTPs will give a beautiful curve and A260:A280 of 1.8!
-bob1-
Another idea would be to use an Agilent Bioanalyzer with a DNA chip. However, the spectrophotometer is much quicker and requires fewer reagents.
-AlyssaK-