Patent ID: 8642266

Claim:
A method for performing an RT-PCR for amplifying a target RNA comprising the steps of lysing a biological sample which contains the target RNA with a lysis buffer comprising between 0.05 M and 1 M of a chaotropic agent, diluting the buffer comprising the target RNA to an extent such that said chaotropic agent is present for a subsequent reverse transcription step in a concentration of 10 to 60 mM, without any intermediate purification step, reverse transcribing said target RNA in the presence of a mixture of first strand cDNA synthesis primers into a first strand cDNA, said mixture consisting of primers hybridizing to a poly-A sequence and/or random primers, and subjecting the transcription mixture comprising the transcribed RNA to multiple cycles of a thermocycling protocol and monitoring amplification of said first strand cDNA in real time, wherein said biological sample consists of not more than 10 cells and the chaotropic agent is guanidine thiocyanate.