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US 9,328,382 B2Granted
Multiple tagging of individual long DNA fragments
AssigneeComplete Genomics, Inc.
InventorsRadoje Drmanac, Brock A. Peters, Andrei Alexeev
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Abstract
This disclosure provides methods and compositions for tagging long fragments of a target nucleic acid for sequencing and analyzing the resulting sequence information in order to reduce errors and perform haplotype phasing, for example.
Claims13
A method of generating tagged sequence reads comprising:
- (a) combining:
- (i) a plurality of long fragments of a target nucleic acid, wherein the fragments are from about 5 kilobases to about 750 kilobases in length;
- (ii) a population of beads, wherein each bead comprises multiple copies of a single polynucleotide comprising a tag-containing sequence, wherein said tag-containing sequence comprises a tag sequence and a transposon end sequence, and wherein different beads comprise different tag sequences;
- and
- (iii) a transposase,
- in a single reaction vessel under conditions in which, for a plurality of beads, the tag sequence associated with an individual bead is transposed into multiple sites of an individual long fragment;
- (b) producing a plurality of subfragments of the long fragments;
- and,
- (c) sequencing subfragments from (b) to produce a plurality of sequence reads including tag sequences.
6 more elements
Dependencies are inferred from the claim text.