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A high-throughput multi-site SNP detection kit and detection method thereof

A detection kit and multi-site technology, applied in biochemical equipment and methods, microbial measurement/inspection, etc., can solve the problems of low detection efficiency, low detection accuracy, low detection throughput, etc., and achieve improved size selectivity , Optimizing the effect of DNA purification magnetic bead components and purification process

Active Publication Date: 2022-07-29
IPE BIOTECHNOLOGY CO LTD
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Problems solved by technology

[0004] For this reason, the embodiment of the present invention provides a high-throughput multi-site SNP detection kit and its detection method to solve the problems of low detection efficiency, low detection throughput, and low detection accuracy in the prior art

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  • A high-throughput multi-site SNP detection kit and detection method thereof
  • A high-throughput multi-site SNP detection kit and detection method thereof
  • A high-throughput multi-site SNP detection kit and detection method thereof

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Embodiment 1

[0048] Example 1. The detection kit of the present invention detects SNP sites of 52 populations in 29 samples

[0049] 1. Experimental materials: Reagents: The high-throughput multi-locus human single nucleotide polymorphism (SNP) detection kit according to the embodiment of the present invention includes: 1) a library preparation kit, which contains 64 sets of labels marked by different sample labels The multiplex PCR primer composition, PCR amplification enzyme, PCR reaction buffer, 2800, DNA purification magnetic beads (purchased from Beckman AMpure); 2) Sequencing template preparation kit (purchased from IonTorrent); 3) Sequencing kit (purchased from IonTorrent) from IonTorrent Corporation). Samples: 29 whole blood samples from Indonesia, Myanmar, the Philippines, Hong Kong, Pakistan, Kyrgyzstan, Tajikistan, Kazakhstan, Nigeria, Zimbabwe, Ukraine and many countries and regions.

[0050] 2. Experimental steps: 1. Library preparation: sample preparation, using chelex-100 m...

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Abstract

The embodiment of the present invention discloses a high-throughput multi-site SNP detection kit and a detection method thereof. The detection kit includes a primer composition consisting of 188 SNP site-specific primer pairs, and the primer composition The nucleotide sequence is shown in SEQ ID NO: 1-SEQ ID NO: 362; the 5' end of each specific primer is connected with a sample tag sequence, and the 5' of the sample tag sequence of the upstream specific specific primer is The end is connected with a sequencing adapter, and the 5' end of the sample tag sequence of the downstream specific primer is connected with a fixed adapter. The detection kit of the embodiment of the present invention can realize a multi-sample, multi-SNP site parallel and stable test kit. The amount of DNA template required for a single reaction system can be less than 1 nanogram, which is much lower than the amount of sample DNA required for mass spectrometry. One measurement can achieve hundreds of SNP sites for dozens of people. The detection room allows large-scale DNA database construction and use .

Description

technical field [0001] Embodiments of the present invention relate to the technical field of biological detection, in particular to a high-throughput multi-site SNP detection kit and a detection method thereof. Background technique [0002] At present, the main methods for typing multiple SNP loci are mass spectrometry and sequencing. Mass spectrometry is mainly matrix-assisted laser desorption ionization time-of-flight mass spectrometry. A piece of DNA containing SNP sites is amplified by PCR. The length of the DNA fragment is about 100-150bp, and then the remaining deoxyribonucleosides in the PCR system are removed by SAP enzyme. Triphosphates (dNTPs) and primers, and then add a single-base extension primer whose 3'-terminal base is close to the SNP site, and four ddNTPs are used instead of dNTPs. In this way, the probe extends only one base at the SNP site, and the attached ddNTP corresponds to the allele at the SNP site. Matrix-assisted laser desorption ionization time...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6869
CPCC12Q1/6869C12Q2531/113C12Q2537/143C12Q2563/159C12Q2535/122
Inventor 焦章平周骋潘雅姣王颖希曲保旺赵怡薛卢艳
Owner IPE BIOTECHNOLOGY CO LTD
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