Next generation sequencing typing reagent kit of 43 STR sites and method

A typing method and next-generation technology, applied in the field of molecular biology, can solve the problems of limited NGS-STR typing technology

Pending Publication Date: 2019-04-02
HEBEI MEDICAL UNIVERSITY
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] Aiming at the technical problems that existing detection methods require the joint application of multiple kits, and the Tm values ​​of primers are difficult to be similar, which limits the NGS-STR typing technology, the present invention provides a next-generation sequencing typing of 43 STR sites Reagent test kit

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  • Next generation sequencing typing reagent kit of 43 STR sites and method
  • Next generation sequencing typing reagent kit of 43 STR sites and method
  • Next generation sequencing typing reagent kit of 43 STR sites and method

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Embodiment 1

[0025] The embodiment of the present invention provides a next-generation sequencing typing kit for 43 STR sites, including next-generation sequencing custom panel kit, blood DNA extraction kit, Qubit dsDNA HS quantitative kit, next-generation sequencing custom panel reagent Kits, real-time fluorescent quantitative kits, labcip quality inspection and analysis kits, and MiSeq on-machine sequencing reagents. The next-generation sequencing custom panel kit includes 79 single-end specific primers for 43 STR sites (the nucleotide sequence is shown in SEQ ID NO.1~SEQ ID NO.79), as well as enzyme digestion reaction buffer, FERA Solution, enzyme mixture in enzyme digestion reaction, ligation reaction buffer, ligation adapter, DNA adapter, ligation solution, targeted PCR reaction buffer, and the 79 PCR amplification single-end specific primers constitute the front and back of conventional PCR primers, Taq DNA polymerase and conventional PCR reaction buffer.

Embodiment 2

[0027] The embodiment of the present invention provides a next-generation sequencing typing method for 43 STR loci. The method uses the next-generation sequencing typing kit for 43 STR loci in Example 1 for sample detection. Including the following steps:

[0028] 1. Extract the DNA of the blood to be tested

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Abstract

The invention relates to a next generation sequencing typing reagent kit of 43 STR sites and a method. The reagent kit comprises a PCR amplification reagent kit, wherein the PCR amplification reagentkit comprises 79 PCR amplification single-end specific primers of the 43 STR sites, and the nucleotide sequence of the PCR amplification single-end specific primers is as shown in SEQ ID NO.1-SEQ ID NO.79. The primers in the reagent kit can enable the polymorphism of each gene locus to be increased, so that probability of exclusion and system effectiveness can be improved, and the testing result is accurate and reliable; and testing results can provide new data for development of a NGS-STR typing technique, and a new detection scheme is provided for cases of target missing person survey, complex genetic relationship identification and the like through united application of a plurality of STR sites of the reagent kits in the prior art.

Description

technical field [0001] The invention relates to the technical field of molecular biology, in particular to a next-generation sequencing typing kit and method for 43 STR sites. Background technique [0002] Paternity testing and personal identification are two main research contents in forensic genetics, and DNA typing technology is the main tool for forensic genetics research. Autosomal short tandem repeats (short tandem repeats, STRs) are the mainstream genetic markers in forensic DNA laboratories, and are the foundation of most forensic DNA databases in countries around the world. At present, the main technology for detecting STR in forensic DNA laboratories is still capillary electrophoresis (CE) technology, which uses multi-color fluorescent labeling multiplex amplification technology to simultaneously type multiple STR loci, and conduct fragment length analysis through CE , to obtain the length information of the STR loci. However, the traditional CE technique can com...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6869C12Q1/6888
CPCC12Q1/6869C12Q1/6888C12Q2600/156C12Q2525/151C12Q2531/113
Inventor 李淑瑾丛斌刘青霞付丽红
Owner HEBEI MEDICAL UNIVERSITY
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