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DIP rapid amplification detection reagent for fully-integrated micro-fluidic chip and application of DIP rapid amplification detection reagent

A technology for detecting people and sequences, applied in the determination/testing of microorganisms, recombinant DNA technology, DNA/RNA fragments, etc., can solve problems such as population inference systems that have not been reported yet, and achieve convenient storage and transportation, simplified transportation methods, and high performance stable effect

Active Publication Date: 2021-08-31
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

Currently foreign quick detectors mainly include: 200, single-channel ID and ANDETM6C, the above rapid detection instruments are all equipped with STR kits for individual identification, and there is no report on the use of the ethnic group inference system for rapid detection equipment

Method used

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  • DIP rapid amplification detection reagent for fully-integrated micro-fluidic chip and application of DIP rapid amplification detection reagent
  • DIP rapid amplification detection reagent for fully-integrated micro-fluidic chip and application of DIP rapid amplification detection reagent
  • DIP rapid amplification detection reagent for fully-integrated micro-fluidic chip and application of DIP rapid amplification detection reagent

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Experimental program
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Embodiment 1

[0088] Embodiment 1, the design of primers and the construction of AIDIP38 chip amplification system

[0089] 1. Primer design and synthesis

[0090] The present invention refers to related literature reports and dbSNP library, and obtains 38 InDel sites through screening (the information of each site is shown in Table 1). Primer premier 5.0 was used to design and optimize primer sequences based on the 38 InDel sites obtained from the above screening, as well as 1 sex site AMEL and 1 site DYS439 on the Y chromosome, and FAM, HEX, FRET-TAMRA and FRET- ROX is fluorescein-labeled at the 5' end of forward primer F at each site. Finally, the primer sequences designed for each locus were obtained, and all primer sequences were synthesized and labeled by Suzhou Xinhai Biotechnology Co., Ltd. See Table 2 for specific information on primer sequences and fluorescent markers, and see figure 1 .

[0091] Table 1. Information of each site

[0092]

[0093]

[0094] Table 2. Prim...

Embodiment 2

[0133] Embodiment 2, the application of AIDIP38 amplification reagent

[0134] 1. Experimental materials and methods

[0135] 1. Experimental materials

[0136] 1) Experimental samples

[0137] Volunteers provided 2 blood card samples, 2 saliva card samples, and 4 oral swab samples. The specific preparation method is as follows:

[0138] Oral swab samples: Scrape the left and right buccal mucosa of volunteers 8 times with the swab.

[0139] 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.

[0140] 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.

[0141] DNA standard 9947A (initial concentration: 10 ng / μL) is a product of Thermo Fisher Scientific, USA.

[0142] 2) Experimental equipment

[01...

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Abstract

The invention discloses a DIP rapid amplification detection reagent for a fully-integrated micro-fluidic chip and application of the DIP rapid amplification detection reagent. According to the invention, a set of DIP rapid amplification reagent which contains 40 loci and can be used for a fully integrated micro-fluidic chip is constructed based on the first fully integrated DNA on-site rapid inspection instrument Quick TargSeq which is independently researched and developed in China, and the reagent can be used for ethnic group inference and can also be used for individual identification. Experiments prove that the amplification reagent can be stored in a micro-fluidic chip card box for a long time in a freeze-drying mode, the whole process of cell lysis, PCR amplification and electrophoretic separation can be completed on samples such as oral swabs, blood cards and saliva cards within 90 min, 'sample input-result output 'type rapid and automatic InDel typing is achieved, and compared with a traditional laboratory method, the result is accurate and reliable, the operation steps are simplified, the pollution risk is reduced, and investigation clues can be provided for cases through rapid and integrated DNA detection of unknown samples.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a DIP rapid amplification detection reagent for a fully integrated microfluidic chip and an application thereof. Background technique [0002] Short tandem repeat (STR) genetic markers commonly used in forensic DNA testing can realize individual identification, but the mutation rate of STR is high, the detection fragment is long, and it cannot detect degraded samples. At the same time, there is no target for STR typing. In some cases where there are no suspects and no other clues, STR typing will not work. Insertion-deletion polymorphism (insertion-deletion polymorphism, InDel) is widespread in the genome, because it combines the favorable characteristics of SNPs and STRs: low mutation rate and quite stable, showing biallelic genes, still belonging to Length polymorphic genetic markers can be detected by conventional PCR-CE. With this genetic marker, degraded DNA can be ...

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 李彩霞韩俊萍赵蕾
Owner INST OF FORENSIC SCI OF MIN OF PUBLIC SECURITY
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