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Nucleic acid adaptors and uses thereof

  • US 9,540,637 B2
  • Filed: 09/29/2011
  • Issued: 01/10/2017
  • Est. Priority Date: 01/09/2008
  • Status: Active Grant
First Claim
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1. A method for circularizing a polynucleotide, comprising:

  • a) providing (i) a linear target polynucleotide having a first end and a second end, (ii) a first double-stranded nucleic acid adaptor with a target joining end and a first overhang end wherein the first overhang end comprises a first overhang nucleic acid sequence, (iii) and a second double-stranded nucleic acid adaptor with a target-joining end and a second overhang end wherein the second overhang end comprises a second overhang nucleic acid sequence, wherein the first and second overhang nucleic acid sequences being at least partially complementary to each other, wherein the first overhang is hybridized to a first single-stranded blocking oligonucleotide over at least a portion of its length and the second overhang is hybridized to a second single-stranded blocking oligonucleotide over at least a portion of its length, and wherein the target-joining end of the first double-stranded nucleic acid adaptor is joined to the first end of the linear target polynucleotide and the target joining end of the second double-stranded nucleic acid adaptor is joined to the second end of the linear target polynucleotide;

    b) removing the first and the second single-stranded blocking oligonucleotides from the first and the second overhangs; and

    c) hybridizing the first overhang having a first nucleic acid sequence with the second overhang having a second nucleic acid sequence, thereby circularizing the polynucleotide, wherein the first and second overhangs are not enzymatically ligated together.

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