Double-label joint design method and preparation method thereof

A technology of tags and adapters, which is applied in the field of design and preparation of double-labeled adapters, can solve problems such as incompatibility with single-end sequencing, increased sequencing time, and increased sequencing costs, so as to avoid false positives, reduce the number of additions, and reduce sequencing costs. Effect

Pending Publication Date: 2020-10-09
ENVELOPE HEALTH BIOTECHNOLOGY CO LTD
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AI Technical Summary

Problems solved by technology

[0006] The existing two-sample label design scheme has the following disadvantages: 1) In order to realize the data acquisition of the double-sample label, it is necessary to add index sequencing primers twice, which increases the sequencing cost; 2) because the sequencing templates of the two sample labels are There are two strands of the DNA library, so the synthesis of the template strand is required before the second sample index sequencing, resulting in increased sequencing time; 3) The current double-sample index design scheme is not compatible with single-end sequencing

Method used

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  • Double-label joint design method and preparation method thereof
  • Double-label joint design method and preparation method thereof
  • Double-label joint design method and preparation method thereof

Examples

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Embodiment 1

[0055] Example 1. Design of dual-sample index adapter and its application to sequencing library construction

[0056] 1. Two-sample index adapter

[0057] 1. Preparation of two-sample index adapter and its library construction kit

[0058] The second sample index in the two-sample index adapter is located between the sequencing primer binding region and the inserted DNA fragment.

[0059] image 3 The structure of the two-sample index adapter shown in A is as follows:

[0060] The double-sample index adapter A is a Y-shaped adapter formed by the annealing of the adapter sequence L-A and the adapter sequence S-A,

[0061] One end of the double-sample index adapter A forms a complementary blunt end or a complementary cohesive end from the complementary region of the adapter sequence L-A and the adapter sequence S-A, and the other end forms a free non-complementary double strand from the non-complementary region of the adapter sequence L-A and the adapter sequence S-A; and the...

Embodiment 2

[0107] Example 2. Design of dual-sample index adapter and its application to sequencing library construction

[0108] 1. Construction of two-sample index adapters

[0109] 1. Construction of double-sample tag adapter sequences

[0110] According to the design scheme of Example 1, design the linker sequence required for constructing the double-sample tag adapter B as shown in Table 1 below;

[0111] The linker sequence in the table was synthesized from Beijing Liuhe Huada Gene Technology Co., Ltd., the purification method was C18 DSL, and the order quantity was 5OD.

[0112] Table 1 Sequences and amplification primer information required to construct adapters

[0113]

[0114] After adapter sequences are ordered, dissolve the master solution to 100 μM in TE buffer.

[0115] 2. Preparation of two-sample index adapters

[0116] Ad01L and Ad01S were mixed according to the same amount of substances, prepared with TE buffer to make a 25 μM joint mixture, left at room temperat...

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Abstract

The invention discloses a double-label joint design method and a preparation method thereof. The invention provides a kit for constructing a DNA molecular sequencing library to be detected. The kit comprises a double-sample label joint, wherein the double-sample label joint is formed by annealing a joint sequence L and a joint sequence S; wherein one end of the double-sample label joint is used for connecting a DNA molecule to be detected; compared with the prior art, the method has the following advantages: (1) new sample tags are introduced to two ends adjacent to inserted DNA fragments, sothat the adding frequency of sequencing primers is reduced, and the sequencing cost is reduced; 2) under the condition that the sequencing cost is not increased, a single-ended sequencing project canrealize double sample labels, so that the false positive problem caused by sample label crosstalk is avoided, and the joint design scheme can realize double sample labels by single-ended sequencing and can realize filtering of wrong sequencing data generated by sample label crosstalk.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a design method and a preparation method of a double-label joint. Background technique [0002] At present, high-throughput sequencing technology has become an important genetic detection technology, which is widely used in scientific research, medical detection, agricultural breeding and judicial identification and other fields. At present, mainstream high-throughput sequencing technology providers include Illumina, Thermo Fisher, Pacbio in the United States, nanopore in the United Kingdom, and BGI in China. In order to reduce the average sequencing cost of samples, in most cases, a strategy of mixing and sequencing multiple sample libraries will be adopted. During the library construction process, a sample index (index) is added to each sample, and the sequencing data can be split into each sample according to the sample index, finally achieving high-throughput and low...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6869C40B50/06
CPCC12Q1/6869C40B50/06C12Q2525/191
Inventor 郑建超汪宇盈羊光辉叶明芝
Owner ENVELOPE HEALTH BIOTECHNOLOGY CO LTD
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