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US 11,441,180 B2Granted
Compositions and methods for accurately identifying mutations
AssigneeFred Hutchinson Cancer Center
InventorJason H. Bielas
Life of this patent
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Abstract
The present disclosure provides compositions and methods for accurately detecting mutations by uniquely tagging double stranded nucleic acid molecules with dual cyphers such that sequence data obtained from a sense strand can be linked to sequence data obtained from an anti-sense strand when sequenced, for example, by massively parallel sequencing methods.
Claims15
A method of sequencing double-stranded cell-free DNA (cfDNA) molecules, the method comprising:
- (a) ligating double-stranded adapters to both ends of the double-stranded cfDNA molecules to form adapter-target complexes, wherein (i) the double-stranded adapters each comprise an identifier sequence, (ii) at least two adapters comprise the same identifier sequence and are ligated to different cfDNA molecules, (iii) cfDNA end sequences together with associated identifier sequences form cyphers at both ends of the cfDNA molecules;
- and (iv) individual adapter-target complexes comprise different pairs of cyphers;
- (b) amplifying the adapter-target complexes to produce a plurality of adapter-target amplification products from each of a first strand and a complementary second strand of the adapter-target complexes;
- (c) sequencing the adapter-target amplification products to produce a plurality of first-strand sequencing reads and a plurality of second-strand sequencing reads;
- (d) comparing the first-strand sequencing reads with the second-strand sequencing reads for each of a plurality of the adapter-target complexes;
- and
- (e) generating error-corrected sequences for each of a plurality of the double-stranded cfDNA molecules by distinguishing erroneous nucleotides in one strand that lack a matched base change in the complementary strand.
4 more elements
Dependencies are inferred from the claim text.