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US 9,005,895 B2Granted
Compositions, methods and kits for nucleic acid synthesis and amplification
AssigneeLife Technologies Corporation
InventorsCora L. Woo, Jennifer Berkman, Priscilla W. Yim
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Abstract
The present invention is directed to compositions, methods and kits useful for the synthesis of nucleic acid molecules. More specifically, compositions, methods and kits are provided for the amplification of nucleic acid molecules in a one-step RT-PCR procedure comprising one or more agents used to increase tolerance to PCR inhibitors.
Claims24
A method of performing RT-PCR of a nucleic acid sample comprising:
- mixing a composition comprising:
- at least one active reverse transcriptase;
- at least one active DNA polymerase;
- and
- a combination of PCR inhibitor blocking agents consisting essentially of 0.2% w/v-0.8% w/v fish gelatin and 500 ng/μl to 6000 ng/μl BSA configured to increase tolerance to one or more PCR inhibitors when present in the nucleic acid sample, with:
- a) a nucleic acid sample;
- b) one or more labeled probes;
- c) one or more primers;
- and
- performing a RT-PCR on the nucleic acid sample.
7 more elements
A method for amplifying a nucleic acid molecule, the method comprising:
- mixing a nucleic acid template with a composition comprising one or more reverse transcriptases, one or more DNA polymerases, and a combination of PCR inhibitor blocking agents consisting essentially of 0.2% w/v-0.8% w/v fish gelatin and 500 ng/μl to 6000 ng/μl BSA, to form a reaction mixture, wherein each of the PCR inhibitor blocking agents is present at a concentration sufficient to reduce PCR inhibition by the PCR inhibitors when present;
- and
- incubating the reaction mixture under conditions sufficient to amplify a nucleic acid molecule complementary to all or a portion of the nucleic acid template.
A method for nucleic acid synthesis, the method comprising:
- mixing one or more first nucleic acid molecules with one or more polymerases, and a combination of PCR inhibitor blocking agents consisting essentially of 0.2% w/v-0.8% w/v fish gelatin and 500 ng/μl to 6000 ng/μl BSA;
- and
- incubating the mixture under conditions sufficient to make one or more second nucleic acid molecules complementary to all or a portion of the one or more first nucleic acid molecules.
Dependencies are inferred from the claim text.