Patent ID: 6613518
Filing Date: 2003-09-02
Classification: C12N,C12Q

Abstract:
A method for quantitative measurement of gene expression of target genes comprising:deparaffinizing a formalin-fixed paraffin-embedded biological tissue sample; rehydrating the deparaffinized sample; combining the rehydrated sample with a chaotropic solution having a pH of about 5 and comprising guanidinium isothiocyanate and a reducing agent; homogenizing and heating the sample and chaotropic solution mixture at a temperature of about 95Â° C. for about 30 minutes; recovering the sample's RNA by extraction from the chaotropic solution using a water insoluble organic solvent; purifying the recovered RNA; converting the purified RNA to cDNA by a reverse transcription reaction; subjecting the cDNA to a PCR reaction in a polymerase chain reaction solution that comprises an oligonucleotide probe suitable for amplifying at least a specified sequence, a polymerase and a fluorochrome; measuring the change that occurs in the intensity of fluorescence as a result of the PCR reaction; and determining, on the basis of the change in the intensity of the fluorescence, the quantity of a nucleic acid having a specified sequence present in the sample.