how do you eat yours... - (Sep/29/2006 )

hi there, me again...

how does everybody generally do their cDNA reactions. i have always done it in 50ul with 1ug RNA. i see most people here use 1ug in 20ul, but if your RNA is not very concentrated, its almost impossible to get 1ug in such a small volume?

I always precipitate my RNA and then resuspend to 1 µg per µL and use 2 µg in a 20 µL reaction. It depends on how sensitive your apparatus is and how much you need to dilute you cDNA at the end to get it in the linear range for amplification.