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STR rapid individual recognition amplification reagent for fully-integrated micro-fluidic chip and application thereof

A reagent, the technology of D2S441, is used in the determination/inspection of microorganisms, DNA/RNA fragments, recombinant DNA technology, etc., and can solve the problem that the DNA typing test method cannot meet the actual needs of public security.

Active Publication Date: 2021-07-13
INST OF FORENSIC SCI OF MIN OF PUBLIC SECURITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Conventional DNA typing and testing methods can no longer meet the actual needs of public security, and it is urgent to realize innovation in the direction of rapid on-site and convenient integration

Method used

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  • STR rapid individual recognition amplification reagent for fully-integrated micro-fluidic chip and application thereof
  • STR rapid individual recognition amplification reagent for fully-integrated micro-fluidic chip and application thereof
  • STR rapid individual recognition amplification reagent for fully-integrated micro-fluidic chip and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0066] Embodiment 1, the design of primers and the construction of RTyper 21 chip detection system

[0067] 1. Primer design and synthesis

[0068] The RTyper 21 chip detection system of the present invention contains 20 CODIS STR loci and 1 Amelogenin sex locus. Design and optimize amplification primers according to each locus (amplification fragment length ranges from 84 to 422bp), and use FAM, HEX, TAMRA, ROX for fluorescent labeling, and all fluorescent labels are in the forward primer F of each site Label the 5' end. The final primer sequences and fluorescent marker specific information designed for each locus are shown in Table 1. All primers were synthesized by Suzhou Xinhai Biotechnology Co., Ltd.

[0069] Table 1. Primer information table

[0070]

[0071]

[0072] 2. Construction of RTyper21 chip amplification detection system

[0073] Based on Quick TargSeq, the first domestic self-developed fully integrated on-site DNA rapid testing instrument, the prese...

Embodiment 2

[0107] Embodiment 2, the application of RTyper21 amplification reagent

[0108] 1. Experimental materials and methods

[0109] 1. Experimental materials

[0110] 1) Experimental samples

[0111] Oral swab samples, blood card samples and saliva card samples provided by volunteers. The three samples were prepared as follows:

[0112] Oral swab samples: Volunteers used sampling swabs to scrape the left and right sides of the inner wall of the oral cavity 8 times.

[0113] Blood card sample: Take 2 drops of blood from the fingertips of volunteers, each drop is about 1cm in diameter, and drop it in the marked circle of the blood collection card.

[0114] Saliva card sample: Take the mucous membrane swabs of the left and right cheeks of the volunteers and transfer them to the saliva card. The above samples are collected and dried in the shade for later use.

[0115] DNA standard 9947A (initial concentration: 10 ng / μL) is a product of Thermo Fisher Scientific, USA.

[0116] 2) ...

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PUM

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Abstract

The invention discloses an STR rapid individual recognition amplification reagent for a fully-integrated micro-fluidic chip and application thereof. According to the invention, a set of micro-fluidic chip rapid amplification reagent-RTyper21 containing 20 CODIS STR gene loci and one Amelogenin sex gene loci is constructed on the basis of a fully integrated DNA on-site rapid inspection instrument Quick TargSeq which is independently researched and developed in China, and a freeze-drying mode is adopted to construct the RTyper21 chip freeze-drying amplification reagent. Experiments prove that the RTyper21 chip freeze-dried amplification reagent can be stored in a micro-fluidic chip card box for a long time, can be used for detecting oral swab, blood card and saliva card samples, can complete the whole process of sample splitting decomposition, PCR amplification and electrophoretic separation within 2 hours, and meets the urgent needs of public security actual combat sites and rapid detection.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to an STR rapid individual recognition amplification reagent for a fully integrated microfluidic chip and an application thereof. Background technique [0002] The use of PCR-STR typing technology can carry out individual identification on the biological evidence of the crime scene, so as to identify or exclude suspects. The routine STR typing process includes DNA extraction, PCR amplification, electrophoretic separation, and typing map analysis. The entire detection process takes approximately 6 to 8 hours. It is time-consuming, has many steps, and complicated operations. It requires high professional skills for operators and is prone to the risk of sample contamination. At present, some major and sudden cases often need to obtain inspection results in a short time to provide direction for case investigation. Conventional DNA typing and testing methods can no longer meet ...

Claims

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Application Information

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IPC IPC(8): C12Q1/6888C12Q1/6858C12N15/11
CPCC12Q1/6888C12Q1/6858C12Q2600/156C12Q2531/113C12Q2565/629
Inventor 李彩霞韩俊萍叶健
Owner INST OF FORENSIC SCI OF MIN OF PUBLIC SECURITY
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