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US 2020/0048707 A1Published application
Compositions and methods for accurately identifying mutations
AssigneeFred Hutchinson Cancer Research 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.
Claims12
-38. (canceled)
A method for reducing an error rate in sequence reads of a double-stranded target nucleic acid molecule, comprising:
- a) ligating the double-stranded target nucleic acid molecule to a double-stranded cypher to form a cypher-target nucleic acid complex, wherein the double-stranded cypher comprises a random or partially random identifier sequence that alone or in combination with an end of the target nucleic acid molecule uniquely labels the double-stranded target nucleic acid molecule;
- b) amplifying each strand of the cypher-target nucleic acid complex to produce a plurality of cypher-target amplification products from each of a first strand and a complementary second strand of the cypher-target nucleic acid complex;
- (c) sequencing the cypher-target amplification products to produce a plurality of first-strand sequencing reads and a plurality of second-strand sequencing reads;
- and
- (d) comparing the first-strand sequencing reads with the second-strand sequencing reads, and generating an error-corrected sequence of the double-stranded target nucleic acid molecule by distinguishing erroneous nucleotides in one strand that lack a matched base change in the complementary strand.
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Dependencies are inferred from the claim text.