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Small RNA Capture, Detection and Quantification

  • US 20150105275A1
  • Filed: 11/04/2013
  • Published: 04/16/2015
  • Est. Priority Date: 11/02/2012
  • Status: Active Grant
First Claim
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1. A method for detecting a mature small RNA, the method comprising:

  • providing a sample comprising a mature small RNA;

    polyadenylating the 3′

    end of the mature small RNA;

    reverse transcribing the polyadenylated mature small RNA using a universal reverse transcription primer, thereby forming a cDNA of the mature small RNA, wherein the universal reverse transcription primer comprises a poly(T) portion and a tail portion, wherein the tail portion comprises a universal primer portion;

    ligating a universal ligation adaptor to the 3′

    end of the cDNA in the presence of a ligation splint, whereby a cDNA ligation product is formed;

    amplifying the cDNA ligation product using a universal forward and reverse primer pair to form an amplification product; and

    detecting the amplification product.

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