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A library-building kit for high-throughput detection of str genetic markers

A genetic marker and high-throughput technology, applied in the field of library construction kits, can solve problems affecting the number of core repeat sequences, the limited number of loci to be analyzed, and mutual interference of fluorescent markers, so as to save detection costs and reduce costs , polymorphism good effect

Active Publication Date: 2019-12-20
承启医学(深圳)科技有限公司
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  • Abstract
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Problems solved by technology

However, this method also has some drawbacks
Mainly include: (1) Fluorescent markers interfere with each other, limitations of imaging techniques, and the number of analyzed loci is very limited
(2) Shadow band (Stutter peak) interference. During the primer extension process of the PCR reaction, the primer strand or template strand slips, resulting in a base non-pairing loop formed by a repeat unit, that is, the strand slippage mismatch mechanism, and finally produces the Stutter peak. thereby affecting the number of repeats of the true core repeat sequence

Method used

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  • A library-building kit for high-throughput detection of str genetic markers
  • A library-building kit for high-throughput detection of str genetic markers
  • A library-building kit for high-throughput detection of str genetic markers

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0052] Example 1: Forensic kinship identification

[0053] The purpose of this example is to detect the number of repeats in the repeat sequence in the STR, wherein the NGS sequencing scheme is used.

[0054] 1. Collection and processing of oral epithelial cells

[0055] (1) Collect human oral epithelial cells with a buccal swab collection tube.

[0056] (2) Oral epithelial cell DNA was extracted with oral swab genome extraction kit (DP322).

[0057] (3) Measure OD260nm and OD280nm with Nanodrop 2000 spectrophotometer, confirm its high purity, and measure its concentration.

[0058] 2. Primer Design for Multiplex PCR Reaction

[0059] (1) The primers are designed as fusion primers, including the specific primer sequence of the target fragment and the sequencing linker sequence, which contains the index sequence.

[0060] (2) The structure of the fusion primer is:

[0061] Upstream primer: 5'-CCTCTCTATGGGCAGTCGGTGAT-3'+ target fragment-specific forward primer;

[0062] Do...

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Abstract

The invention provides a library building kit for high flux detection of STR genetic markers. The kit comprises multiple pairs of fusion primers for amplifying target STR loci, each pair of fusion primer respectively aims at different STR loci on a DNA template, and each pair of fusion primer comprises a sequencing joint sequence and a specific primer sequence which aims at the STR locus from a 5'end to a 3'end in order. The kit can be used for realizing construction of a STR locus amplicon sequencing library by one-time amplification, the operation process of library building is simplified, time and cost for building the library are reduced, and sequencing quality is not influenced.

Description

technical field [0001] The invention belongs to the field of forensic genetics, and relates to a library construction kit for using high-throughput sequencing technology for forensic kinship identification and analyzing autosomal STR genetic markers based on NGS sequencing. Background technique [0002] Forensic DNA detection technology is to carry out individual identification and paternity identification in forensic medicine by testing the sequence polymorphism and length polymorphism of genetic material DNA. Today, this technology has played a significant role in investigating and solving crimes, finding the source of trafficked children and unknown corpses, identifying individuals in death cases, and confirming crimes. It has become a routine technology for forensic evidence inspection and provides a powerful weapon for combating crimes. . [0003] The research and analysis object of forensic DNA testing technology is mainly the polymorphism of DNA in organisms. Human ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6888C40B50/06
CPCC12Q1/6806C12Q1/6869C40B50/06C12Q2531/113C12Q2537/143C12Q2525/191C12Q2525/151C12Q2535/122
Inventor 王旭钟嘉泳张弓董鸣余卓
Owner 承启医学(深圳)科技有限公司
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