Forgot to stop reverse transcription reaction - (Jun/25/2010 )

Hi All,

I wanted to find out what the consequences are if you don't end the reverse transctiption reaction by heating at 95oC, but rather immediatly chill the reaction after the reverse transcription step?

Cheers,
Massiveattack

It is not too late to do that now.

Thanks for the reply! I used the monsterscript kit and performed the reverse transcription at 60oC for 40mins, then instead of doinf the 95oC step I paced the reactions on ice and then stored at -20oC. Can I leave the reactions at -20oC for now and then denature/inactivate the RT protein at any time before I use it for qPCR or is important to do this immediately? Also, what would happen if the RT protein is active when qPCR is performed? Wouldn't it be inactivated in the initial denaturation step in the qPCR anyway?

Cheers,
massiveattack

it will be inactivated in the first denaturation step in the qPCR, so don't worry.

Hi,

Thanks for the reply. I thought that this would be the case.

Cheers,
massiveattack