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US 2024/0200131 A1Published application
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.
Claims17
-38. (canceled)
A method for sequencing double-stranded target DNA molecules, comprising:
- (a) obtaining double-stranded target DNA molecules from a biological sample and end-repairing double-stranded target DNA molecule fragments;
- (b) ligating adapter molecules to both ends of a plurality of original double-stranded target DNA molecule fragments to form a population of adapter-target DNA complexes;
- wherein the adapter molecules each comprise an identifier sequence to uniquely label the double-stranded target DNA molecule fragments, such that ligated double-stranded target DNA molecule fragments in the population of adapter-target DNA complexes are distinguishable from one another by associated identifier sequences alone or in combination with an associated end sequence of a target DNA molecule fragment;
- and
- (c) for each of a plurality of the adapter-target DNA complexes, amplifying each strand of the adapter-target DNA complex to produce a plurality of first strand adapter-target DNA complex amplicons and a plurality of second strand adapter-target DNA complex amplicons;
- wherein the plurality of first strand adapter-target DNA complex amplicons and the plurality of second strand adapter-target DNA complex amplicons derived from the same original double-stranded target DNA molecule fragment are relatable by the associated identifier sequences alone or in combination with the associated end sequence of the target DNA molecule fragment.
3 more elements
Dependencies are inferred from the claim text.