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US 11,597,973 B2Granted
Compositions and methods for detecting rare sequence variants
AssigneeAccuragen Holdings Limited
InventorPaul Ling-Fung Tang
Life of this patent
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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.
Claims28
A method for processing or analyzing a biological sample of a subject, comprising:
- (a) obtaining said biological sample of said subject;
- (b) using a ribonucleic acid (RNA) molecule or derivative thereof derived from said biological sample to generate a circular RNA molecule;
- (c) subjecting said circular RNA molecule or derivative thereof to reverse transcription to generate a complementary deoxyribonucleic acid (cDNA) concatemer;
- (d) creating an amplicon of a target sequence of said cDNA molecule using linear amplification to amplify said cDNA concatemer or derivative thereof using a primer that comprises (i) a sequence that binds to said target sequence or a sequence that binds to a complement of a junction sequence between a 5′ end and a 3′ end of said RNA molecule in said circular RNA molecule comprising said target sequence;
- and (ii) an adaptor sequence;
- and
- (e) subjecting said amplicon of said cDNA concatemer or derivative thereof to sequencing to identify a sequence of said RNA molecule.
4 more elements
Dependencies are inferred from the claim text.