Differential tagging of RNA for preparation of a cell-free DNA/RNA sequencing library
First Claim
Patent Images
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.
3 Assignments
0 Petitions
Accused Products
Abstract
In various aspects, the present disclosure provides methods, compositions, reactions mixtures, kits, and systems for sequencing both RNA and DNA from a single source sample. In some embodiments, RNA is treated so as to differentiate RNA sequences from DNA sequences derived from the same sample. In some embodiments, the RNA and DNA are cell-free polynucleotides.
23 Citations
14 Claims
-
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. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14)
-
Specification