IMPROVED LYSIS AND REVERSE TRANSCRIPTION FOR MRNA QUANTIFICATION

摘要

The present invention is directed to a method for performing an RT-PCR for amplifying a target RNA comprising the steps of (i) cultivation of a population of adherent cells in a cell culture vessel (ii) lysis of said population of adherent cells which is supposed to contain said target RNA in said sample vessel with a lysis buffer comprising between 0.05 M and 1 M of a chaotropic agent (iii) adding reagents to said sample vessel which are necessary to perform a reverse transcription reaction such that the said chaotropic agent is present in a concentration of about 10 to 60 mM in said sample vessel, and reverse transcribing said target RNA and (iv) amplifying said first strand cDNA by means of subjecting said sample to multiple cycles of a thermocycling protocol.