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Methods of RNA amplification in the presence of DNA

  • US 7,846,666 B2
  • Filed: 03/20/2009
  • Issued: 12/07/2010
  • Est. Priority Date: 03/21/2008
  • Status: Expired due to Fees
First Claim
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1. A method of selectively generating multiple copies of a polynucleotide sequence of or complementary to target RNA which is in a sample comprising DNA, said method comprising the steps of:

  • (a) hybridizing to the target RNA in said sample a first primer comprising a sequence (A) that is not complementary to the target RNA, and a sequence (B) at the 3′

    -end which hybridizes to the target RNA;

    (b) extending the first primer with at least one enzyme comprising RNA-dependent DNA polymerase activity in the presence of at least one compound comprising DNA-dependent DNA polymerase inhibitor activity, whereby DNA-dependent DNA polymerase activity of the enzyme comprising RNA-dependent DNA polymerase activity is inhibited by said compound, whereby a complex comprising a primer extension product and the target RNA is produced, whereby the first primer extension product comprises a sequence (Y) that is complementary to the target RNA and comprises sequence (A);

    (c) disabling or removing at least one compound comprising DNA-dependent DNA polymerase inhibitor activity; and

    (d) producing multiple copies of a polynucleotide sequence complementary to the target RNA and/or complementary to sequence (Y) using sequence (A).

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