Compositions and methods for detecting rare sequence variants
AssigneeAccuragen Holdings Limited
InventorsShengrong LIN, Zhaohui SUN, Grace Qizhi ZHAO, Paul Ling-Fung TANG
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Abstract
In some aspects, the present disclosure provides methods for identifying sequence variants in a nucleic acid sample. In some embodiments, a method comprises identifying sequence differences between sequencing reads and a reference sequence, and calling a sequence difference that occurs in at least two different circular polynucleotides, such as two circular polynucleotides having different junctions, as the sequence variant. In some aspects, the present disclosure provides compositions and systems useful in the described methods.
Claims21
-52. (canceled)
A method of distinguishing a sequence variant from an error in a cell-free polynucleotide, comprising:
- (a) circularizing said cell-free polynucleotide or a derivative thereof to form a circular polynucleotide, wherein said cell-free polynucleotide is not enzymatically phosphorylated prior to circularization;
- (b) amplifying said circular polynucleotide of (a) to produce a plurality of concatemers, wherein a concatemer of said plurality of concatemers comprises at least two copies of a sequence of said circular polynucleotide;
- (c) sequencing said plurality of concatemers to produce a plurality of sequencing reads;
- and
- (d) identifying a sequence difference between a sequencing read of said plurality of sequencing reads and a reference sequence as said sequence variant only when said sequence difference occurs in at least two copies of said circular polynucleotide in said concatemer and not when said sequence difference occurs in only one copy of said circular polynucleotide in said concatemer, thereby distinguishing said sequence variant from said error.
Dependencies are inferred from the claim text.