Composition and methods for rt-pcr comprising an anionic polymer
AssigneeQiagen GmbH
InventorsNan Fang, Andreas Missel
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Abstract
The present invention is in the fields of molecular biology. The present invention is directed to novel compositions, methods and kits useful for the generation of nucleic acids from an RNA template and further nucleic acid replication. Specifically, the invention is directed to the generation and amplification of nucleic acids by reverse transcriptase-polymerase chain reaction.
Claims16
A composition suitable for the generation of nucleic acids from an RNA template, said composition comprising
- an anionic polymer that is not a nucleic acid,
- one or more oligonucleotide primers,
- one or more nucleotides or derivatives thereof, and
- two or more different polymerases, at least one of which has reverse transcriptase activity and at least one of which has nucleic acid polymerase activity.
A method for generating a nucleic acid from an RNA template comprising the steps:
- a) adding said RNA template to a reaction mixture, said reaction mixture comprising at least one reverse transcriptase, at least one nucleic acid polymerase, an anionic polymer that is not a nucleic acid, and one or more oligonucleotide primers;
- and
- b) incubating said reaction mixture under conditions sufficient to allow polymerization of a nucleic acid molecule complementary to a portion of said RNA template.
A kit for performing coupled RT-PCR, said kit comprising at least one reverse transcriptase, at least one DNA polymerase, and at least one anionic polymer that is not a nucleic acid.
Dependencies are inferred from the claim text.