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Methods for detecting small RNA species

  • US 20080241831A1
  • Filed: 03/28/2007
  • Published: 10/02/2008
  • Est. Priority Date: 03/28/2007
  • Status: Abandoned Application
First Claim
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1. A method for determining the presence of a small target ribonucleotide sequence in a sample, said method comprising(a) modifying ribonucleotide species contained in the sample by polyadenylating the 3′

  • ends;

    (b) converting the polyadenylated ribonucleotide species into a plurality of complementary DNA (cDNA) sequences;

    (c) immobilizing said plurality of cDNA sequences to a solid support;

    (d) contacting said immobilized cDNA species with a pool of probe nucleic acids under conditions that allow sequence specific annealing, wherein each probe nucleic acid corresponds to a small target ribonucleotide sequence;

    (e) extending said probe nucleic acids in a manner complementary to the immobilized cDNA species;

    (f) removing the extended probe nucleic acids from the immobilized cDNA species;

    (g) amplifying the extended probe nucleic acids to generate amplicons, and(h) detecting said amplicons, wherein detection of each amplicon indicates the presence of a small target ribonucleotide sequence.

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