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Methods, compositions and kits for the improved detection of small RNA molecules

  • US 8,980,558 B2
  • Filed: 12/05/2013
  • Issued: 03/17/2015
  • Est. Priority Date: 10/09/2007
  • Status: Active Grant
First Claim
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1. A method for amplifying a small RNA target sequence less than about 100 nucleotides in length, which comprises:

  • (a) contacting a sample comprising the small RNA target sequence with;

    (i) a primer that is sufficiently complementary to the small RNA target sequence to hybridize thereto, and(ii) an RNA-dependent DNA polymerase having reverse transcriptase activity;

    (b) synthesizing cDNA complementary to the small RNA target sequence in the presence of a mixture of dNTPs comprising at least one base-modified, duplex-stabilizing dNTP that is a substrate for the RNA-dependent DNA polymerase; and

    (c) performing a nucleic acid amplification reaction using amplification primers that are sufficiently complementary to the small RNA target sequence and its complement to hybridize thereto and produce amplification products in the presence of DNA polymerase.

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