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Method of synthesizing cdna

  • US 20060246453A1
  • Filed: 03/29/2004
  • Published: 11/02/2006
  • Est. Priority Date: 03/28/2003
  • Status: Active Grant
First Claim
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1. A method for synthesizing cDNA possessing a consecutive sequence starting with a nucleotide adjacent to a cap structure of mRNA, which method comprises the steps of:

  • (i) annealing a double-stranded DNA primer and an RNA mixture containing mRNA possessing a cap structure, (ii) preparing a conjugate of an mRNA/cDNA heteroduplex and a double-stranded DNA primer by synthesizing the first-strand cDNA primed with the double-stranded DNA primer using reverse transcriptase, and (iii) circularizing the conjugate of the mRNA/cDNA heteroduplex and the double-stranded DNA primer by joining the 3′ and

    5′

    ends of the DNA strand containing cDNA using ligase.

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