ask for help about the RT-PCR - does different aoumnt of RNA affect the RT efficiency? (Jan/05/2007 )
does different aoumnt of RNA affect the RT efficiency?
I got the lower amount (nanogram/ul) of RNA samples from Laser micro dissection tissue. and the volume of RNA solution is equivalent. while the concentration is different. So it is hard to adjust the amount of RNA to the same concentration.
Can I input the different amount of RNA into the Reverse transcript reactions? after RT, according to how much RNA i input into the reaction, adjust to the final concentration?
I 'm not sure whether it will influence the realtime PCR.
please help me! thank you
It should not unless you have some impurities in your RNA prep that inhibit the reaction. This can be accouted for by using a housekeeping gene as an endogenous control.