Patent ID: 6183995
Filing Date: 2001-02-06
Classification: C12N,C12Q,Y10T

Abstract:
A method for the production of cDNA representative of complete representational cellular mRNA from a wax-embedded biological sample comprising:(a) extracting RNA from the wax-embedded sample by removing the DNA in the biological sample with a DNAse and without a step to selectively eliminate rRNA by selection for polyadenylated RNA to produce sample cellular RNA; and then(b) synthesizing a first strand of cDNA complementary to the sample cellular mRNA by contacting the sample RNA to one or more oligonucleotides and a reverse transcriptase to create single strand cDNA copies of the cellular RNA representative of the complete cellular mRNA; and(c) ligating a pair of complementary oligonucleotide adaptors to the cDNA wherein the adaptors are of different lengths.