Reduced inhibition of one-step RT-PCR

摘要

The present invention provides a method for amplifying a nucleic acid molecule. The method involves mixing an RNA template with a composition having a reverse transcriptase, a DNA polymerase and a RT inhibition reducer. The RT inhibition reducer can be Sso7d, Sac7d, Sac7e, Sso7e, AluI methylase, suramin, a phosphorothioate oligodeoxycytosine, a phosphorothioate oligodeoxyadenine, a phosphorothioate oligodeoxythymine or poly(rA)(dT). The mixing forms a mixture that is incubated under conditions sufficient to synthesize a DNA molecule complementary to all or a portion of the RNA template, thereby amplifying the nucleic acid molecule.

主权项

1. A method of reverse transcribing a cDNA molecule from an RNA template, comprising:
mixing the RNA template with a composition comprising a reverse transcriptase, a DNA primer, and a phosphorothioate oligodeoxynucleotide, wherein the phosphorothioate oligodeoxynucleotide is present in an amount from 0.01 nM to 25 nM; and incubating the mixture at a temperature sufficient to synthesize a DNA molecule complementary to at least a portion of the RNA template, thereby reverse transcribing the cDNA molecule.