Methods and compositions for nucleic acid sequencing

Patent metadata

Field	Value
Publication number	US-10557133-B2
Application number	US-201314766089-A
Country	US
Kind code	B2
Filing date	Mar 13, 2013
Priority date	Mar 13, 2013
Publication date	Feb 11, 2020
Grant date	Feb 11, 2020

Abstract

Embodiments of the present invention relate to sequencing nucleic acids. In particular, embodiments of the methods and compositions provided herein relate to preparing nucleic acid templates and obtaining sequence data therefrom.

First claim

What is claimed is:

1. A method for preparing a library of template nucleic acids to obtain sequence information from a target nucleic acid, said method comprising:
(a) contacting a template nucleic acid with a plurality of transposomes to generate a tagmented template nucleic acid, wherein at least some of the transposomes each comprise a first transposon sequence, a second transposon sequence noncontiguous with said first transposon sequence, and a transposase associated with the first transposon sequence and the second transposon sequence, wherein a plurality of transposons are inserted into the target nucleic acid;
(b) compartmentalizing the tagmented template nucleic acid comprising said plurality of inserted transposons into each vessel of a plurality of vessels, wherein the compartmentalizing comprises distributing the tagmented template nucleic acid between the plurality of vessels to provide each vessel with an amount of tagmented template nucleic acid less than about one haploid equivalent of the target nucleic acid; and
(c) removing the transposase from the template nucleic acid.

2. The method of claim 1 , wherein step (c) comprises a method selected from the group consisting of adding a detergent, changing temperature, changing pH, adding a protease, adding a chaperone, and adding a polymerase.

3. The method of claim 1 , wherein the first transposon sequence comprises a first primer site and the second transposon sequences comprise a second primer site, wherein the first primer site further comprises a first barcode and the second primer site further comprises a second barcode.

4. The method of claim 3 , wherein the first barcode and second barcode are different.

5. The method of claim 1 , wherein step (a) further comprises enriching the template nucleic acid for a sequence of interest.

6. The method of claim 1 , further comprising obtaining sequence information from the template nucleic acid.

7. The method of claim 6 , wherein the sequence information comprises haplotype sequence information.

8. The method of claim 1 , wherein the template nucleic acid is from a single cell.

9. The method of claim 1 , wherein the template nucleic acid is total genomic DNA.

Assignees

• Illumina Inc

Inventors

• Steemers Frank J
• Amini Sasan
• Gunderson Kevin L
• Pignatelli Natasha
• Goryshin Igor

Classifications

• C12Q1/6806

  Preparing nucleic acids for analysis, e.g. for polymerase chain reaction [PCR] assay (C12Q1/6804 takes precedence) · CPC title

• C12Q1/6869

  Methods for sequencing · CPC title

• C12N15/1065 Primary

  Preparation or screening of tagged libraries, e.g. tagged microorganisms by STM-mutagenesis, tagged polynucleotides, gene tags · CPC title

• C12N15/1075

  by coupling phenotype to genotype, not provided for in other groups of this subclass · CPC title