Dna library sequence for increasing number of chip probes and application thereof

Patent metadata

Field	Value
Publication number	US-2025043273-A1
Application number	US-202118719318-A
Country	US
Kind code	A1
Filing date	Dec 24, 2021
Priority date	Dec 24, 2021
Publication date	Feb 6, 2025
Grant date	—

Abstract

A DNA library sequence for increasing the number of chip probes and an application thereof. The DNA library sequence comprises two or more position information labeling sequences and three or more fixed sequences, wherein the position information labeling sequences and the fixed sequences are arranged at intervals. The DNA library sequence increases the number of probes of a chip used for spatiotemporal omics, and then a capture number of transcript genes is increased. The present invention has a very high application prospect.

First claim

1 . A DNA library sequence for increasing the number of chip probes, which comprises two or more position information labeling sequences and three or more fixed sequences, wherein the position information labeling sequences and the fixed sequences are arranged at intervals.

2 . The DNA library sequence according to claim 1 , wherein the DNA library sequence comprises, in order from a direction of 5′ to 3′: a first fixed sequence, a first position information labeling sequence, a second fixed sequence, a second position information labeling sequence, and a third fixed sequence.

3 . The DNA library sequence according to claim 2 , wherein the position information labeling sequence consists of at least 1 N, and each N is independently A, T, C, or G.

4 . The DNA library sequence according to claim 1 , wherein a first fixed sequence and a last fixed sequence of the DNA library sequence are each connected to an amplification site.

5 . The DNA library sequence according to claim 1 , wherein the DNA library sequence is connected to a nucleic acid molecule comprising a capture sequence capable of capturing nucleic acid in a sample.

6 . The DNA library sequence according to claim 1 , wherein the fixed sequence contains a sequencing primer required for decoding the position information labeling sequence and a sequence that connects position information with cDNA of a subsequent single-cell transcriptome.

7 . A kit, comprising:
(1) the DNA library sequence according to claim 1 ; (2) a reaction mix for amplification.

8 . A method for preparing a chip with an increased number of probes, the method containing:
(1) synthesizing the DNA library sequence according to claim 1 ; (2) amplifying the DNA library sequence; (3) decoding the position information labeling sequence by sequencing; (4) adding a capture sequence.

9 . A chip with an increased number of probes prepared by the method according to claim 8 .

10 . A method for detecting spatial information of a nucleic acid in a sample, wherein the method comprises using the DNA library sequence according to claim 1 .

11 . A method for detecting spatial information of a nucleic acid in a sample, wherein the method comprises using the kit according to claim 7 .

12 . A method for detecting spatial information of a nucleic acid in a sample, comprises using the chip according to claim 9 .

13 . The method according to claim 8 , wherein the amplification is bridging PCR amplification, rolling circle amplification, multiple displacement amplification, or emulsion PCR amplification.

14 . The method according to claim 8 , wherein the sequencing is ligation sequencing or sequencing by synthesis.

15 . The DNA library sequence according to claim 3 , wherein the position information labeling sequence has 5 or more Ns.

16 . The DNA library sequence according to claim 15 , wherein the position information labeling sequence has 5 to 50 Ns.

17 . The DNA library sequence according to claim 2 , wherein the DNA library sequence comprises a third position information labeling sequence and a fourth fixed sequence connected downstream to the third fixed sequence.

18 . The DNA library sequence according to claim 17 , wherein the DNA library sequence comprises up to an nth position information labeling sequence and an (n+1)th fixed sequence; n is an integer greater than 3.

Assignees

• Bgi Shenzhen

Inventors

• Liao Sha
• Chen Ao
• Zhang Wenwei
• Xu Xun
• Chen Xi
• Fu Defeng

Classifications

• C40B40/06

  Libraries containing nucleotides or polynucleotides, or derivatives thereof · CPC title

• C40B20/02

  Identifying library members by their fixed physical location on a support or substrate · CPC title

• C12N15/1065 Primary

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

• C12Q1/6841 Primary

  In situ hybridisation · CPC title