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Differential tagging of RNA for preparation of a cell-free DNA/RNA sequencing library

  • US 10,144,962 B2
  • Filed: 06/29/2017
  • Issued: 12/04/2018
  • Est. Priority Date: 06/30/2016
  • Status: Active Grant
First Claim
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1. A method of distinguishing sequences of RNA and DNA in a sample, the method comprising:

  • a. obtaining a sample comprising both RNA and DNA;

    b. reverse transcribing the RNA to produce cDNA/RNA hybrid molecules;

    c. degrading the RNA of the hybrid molecules to produce single-stranded cDNA;

    d. preferentially joining a tag oligonucleotide comprising a tag sequence to the single-stranded cDNA in a reaction comprising a single-stranded DNA ligase to produce tagged cDNA;

    e. sequencing the DNA and the tagged cDNA; and

    f. using a processor to group RNA-derived sequences separately from DNA-derived sequences based on the presence or absence of the tag sequence, or a complement of the tag sequence;

    wherein the reverse transcribing, preferentially joining, and sequencing are performed in the presence of the DNA.

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