Multiple tagging of individual long DNA fragments
First Claim
Patent Images
1. A method of sequencing a target nucleic acid without the use of nanodrops comprising:
- (a) combining in a single mixture (i) first fragments of the target nucleic acid, and (ii) a population of beads, wherein each bead comprises oligonucleotides immobilized thereon, said oligonucleotides comprising a tag-containing sequence, wherein each tag-containing sequence comprises a tag sequence, wherein the oligonucleotides immobilized on the same individual bead comprise the same tag-containing sequence and a majority of beads have different tag sequences;
(b) introducing into a plurality of the first fragments multiple copies of a tag sequence, wherein the multiple copies are from a single bead per first fragment;
(c) producing a plurality of subfragments from the plurality of first fragments, wherein a plurality of the subfragments comprise one or more tags;
(d) sequencing the plurality of subfragments to produce a plurality of sequence reads.
1 Assignment
0 Petitions
Accused Products
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.
74 Citations
19 Claims
-
1. A method of sequencing a target nucleic acid without the use of nanodrops comprising:
-
(a) combining in a single mixture (i) first fragments of the target nucleic acid, and (ii) a population of beads, wherein each bead comprises oligonucleotides immobilized thereon, said oligonucleotides comprising a tag-containing sequence, wherein each tag-containing sequence comprises a tag sequence, wherein the oligonucleotides immobilized on the same individual bead comprise the same tag-containing sequence and a majority of beads have different tag sequences; (b) introducing into a plurality of the first fragments multiple copies of a tag sequence, wherein the multiple copies are from a single bead per first fragment; (c) producing a plurality of subfragments from the plurality of first fragments, wherein a plurality of the subfragments comprise one or more tags; (d) sequencing the plurality of subfragments to produce a plurality of sequence reads. - View Dependent Claims (2, 3, 4, 5, 6, 7, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19)
-
- 8. The method of 7 wherein the organism is a human.
Specification