US 2003/0113712 A1 · Compositions and methods for reverse transcriptase-polymerase chain reaction (RT-PCR)

Abstract

The present invention is directed to compositions and methods useful for the amplification of nucleic acid molecules by reverse transcriptase-polymerase chain reaction (RT-PCR). Specifically, the invention provides compositions and methods for the amplification of nucleic acid molecules in a simplified one- or two-step RT-PCR procedure using combinations of reverse transcriptase and thermostable DNA polymerase enzymes in conjunction with sulfur-containing molecules or acetate-containing molecules (or combinations of such sulfur-containing molecules and acetate-containing molecules), and optionally bovine serum albumin. The invention thus facilitates the rapid and efficient amplification of nucleic acid molecules and the detection and quantitation of RNA molecules. The invention also is useful in the rapid production and amplification of cDNAs which may be used for a variety of industrial, medical and forensic purposes.

Claims56

Dependent claims

A method for amplifying a nucleic acid molecule, said method comprising

(a) mixing an RNA template with a composition comprising a Moloney murine leukemia virus (M-MLV) reverse transcriptase, one or more DNA polymerases and one or more sulfur-containing molecules at a concentration of at least 18 mM sulfur, to form a mixture;
and
(b) incubating said mixture under conditions sufficient to amplify a DNA molecule complementary to all or a portion of said RNA template.

Dependencies are inferred from the claim text.

Compositions and methods for reverse transcriptase-polymerase chain reaction (RT-PCR)

Life of this patent

Compositions and methods for reverse transcriptase-polymerase chain reaction (RT-PCR)

Life of this patent

Abstract

Claims56