Methods for analyzing nucleic acids from single cells

Patent metadata

Field	Value
Publication number	US-10633702-B2
Application number	US-201916443703-A
Country	US
Kind code	B2
Filing date	Jun 17, 2019
Priority date	Aug 20, 2009
Publication date	Apr 28, 2020
Grant date	Apr 28, 2020

Abstract

Aspects of the present invention include analyzing nucleic acids from single cells using methods that include using tagged polynucleotides containing multiplex identifier sequences.

First claim

What is claimed:

1. A system for multiplexed nucleic acid analysis, comprising:
a solid support having a plurality of oligonucleotide tags disposed thereon, wherein the plurality of oligonucleotide tags is (a) covalently attached to a surface of the solid support and (b) configured to label a plurality of sample polynucleotides from a cell, wherein an oligonucleotide tag of said plurality of oligonucleotide tags comprises:
(i) a first tag sequence configured to distinguish a sample polynucleotide originating from a cell from sample polynucleotides originating from other cells; and
(ii) a second tag sequence configured to distinguish said sample polynucleotide from other sample polynucleotide from the same cell and having the same sequence as said sample polynucleotide.

2. The system of claim 1 , wherein an oligonucleotide tag of said plurality of oligonucleotide tags comprises a capture sequence that is configured to capture said sample polynucleotides.

3. The system of claim 1 , wherein the first tag sequence has a length of 8 to 20 nucleotides.

4. The system of claim 1 , wherein said plurality of oligonucleotide tags comprises 1,000 different first tag sequences, 10,000 different first tag sequences, or 1,000,000 different first tag sequences.

5. The system of claim 4 , wherein said plurality of oligonucleotide tags comprises 1,000 different first tag sequences.

6. The system of claim 5 , wherein said plurality of oligonucleotide tags comprises 10,000 different first tag sequences.

7. The system of claim 6 , wherein said plurality of oligonucleotide tags comprises 1,000,000 different first tag sequences.

8. The system of claim 1 , wherein said plurality of oligonucleotide tags comprises 1,000 different second tag sequences, 10,000 different second tag sequences, or 1,000,000 different second tag sequences.

9. The system of claim 8 , wherein said plurality of oligonucleotide tags comprises 1,000 different second tag sequences.

10. The system of claim 9 , wherein said plurality of oligonucleotide tags comprises 10,000 different second tag sequences.

11. The system of claim 10 , wherein said plurality of oligonucleotide tags comprises 1,000,000 different second tag sequences.

12. The system of claim 1 , wherein the second tag sequence has a length of 8 to 20 nucleotides.

13. The system of claim 1 , wherein an oligonucleotide tag of said plurality of oligonucleotide tags comprises a sequencing adaptor.

14. The system of claim 1 , wherein the solid support comprises a well.

15. The system of claim 1 , wherein the solid support comprises a planar solid phase support.

16. The system of claim 1 , wherein the solid support comprises a glass slide.

Assignees

• 10X Genomics Inc

Inventors

• Brenner Sydney
• Mikawa Gi
• Osborne Robert
• Slatter Andrew

Classifications

• C12N15/1065

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

• C12Q1/6874 Primary

  involving nucleic acid arrays, e.g. sequencing by hybridisation · CPC title

• C12Q1/686

  Polymerase chain reaction [PCR] · CPC title

• C12Q1/6806

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

• C12Q1/6855 Primary

  Ligating adaptors · CPC title