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Fluorescent multiplex amplification system of 37 Y-STR gene locuses and Y-Indel, kit and application thereof

A compound amplification system, y-indel technology, is applied in the determination/inspection of microorganisms, biochemical equipment and methods, DNA/RNA fragments, etc., which can solve the problem of cumulative individual identification and cumulative non-parent exclusion rate, which needs to be further improved, etc. problem, to achieve the effect of high cumulative individual recognition, high individual recognition rate, and strong species specificity

Active Publication Date: 2018-09-14
SUZHOU MICROREAD GENETICS
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the existing kits on the market have a small number of loci, and there is no multiple amplification system that includes these 35 loci at the same time. The cumulative individual recognition ability and cumulative non-parent exclusion rate still need to be further improved

Method used

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  • Fluorescent multiplex amplification system of 37 Y-STR gene locuses and Y-Indel, kit and application thereof
  • Fluorescent multiplex amplification system of 37 Y-STR gene locuses and Y-Indel, kit and application thereof
  • Fluorescent multiplex amplification system of 37 Y-STR gene locuses and Y-Indel, kit and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0046] Example 1 Selection of 37 Y chromosome STR loci and 1 Y-Indel, fluorescent label combination and primer design

[0047] In terms of locus selection, this product combines 20 core loci and 15 preferred loci from the Ministry of Public Security, adds a Y-Indel locus, and also contains a unique locus DYF404S1.

[0048] Divide 37 loci and 1 Y-Indel into five groups. After repeated experiments and verification, determine the optimal fluorescent dye combination scheme, Y-Indel, DYS393, DYS570, DYS19, DYS392, DYS549, Y-GATA-H4 , DYS444 is marked with FAM; DYS460, DYS458, DYS481, DYS635, DYS448, DYS533, DYS596 are marked with HEX; DYS456, DYS389I, DYS390, DYS389II, DYS438, DYS447 and DYS596 are marked with L552; DYS391, DYS439, DYS437, DYS385a / b DYS643, DYS518 are marked with LR600; DYS576, DYF404S1, DYF387S1, DYS627, DYS527a / b, DYS557 are marked with TET-592.

[0049] Design a pair of primers for each of the above loci, and perform amplification experiments with a single pair of pri...

Embodiment 2

[0059] Capillary electrophoresis detection of amplified products

[0060] Before detection, the product is denatured and mixed with the molecular weight internal standard, 1ul product + 0.5ul molecular weight internal standard + 8.5ul HIDI (deionized formamide), mix and centrifuge briefly, and immediately after denaturation at 95°C for 5 minutes, ice bath 10min, use ABI series genetic analyzer for electrophoresis detection, such as figure 1 . It can be seen from the figure that each locus can be typed accurately, with good specificity, no non-specific bands appear, and the peak height balance of each locus is also good.

Embodiment 3

[0062] Species specific experiments

[0063] Amplify the templates of pigs, cattle, sheep, and chickens, and detect the species specificity of 37 Y chromosome STR loci and 1 Y-Indel multiplex amplification system. The results are as follows figure 2 Shown. Through the amplification of other species, no bands were amplified, indicating that the 34 pairs of primers have good species specificity.

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Abstract

The invention provides a fluorescent multiplex amplification system of 37 Y-STR gene locuses and a Y-Indel, a kit and application thereof, belonging to the technical field of biological detection. Thekit can amplify and analyze 37 Y-STR gene locuses and a Y-Indel at the same time. Six colors of fluorescence are used for marking, the size of the amplified product is 70-490bp, the operation is simple, and very high cumulative individual discernment capability and cumulative probability of paternity exclusion are realized.

Description

Technical field [0001] The present invention belongs to the technical field of biological detection and is used to detect the polymorphism of short tandem repeat loci in the human genome. Specifically, the present invention relates to a fluorescent compound amplification system of 37 Y chromosome STR loci and 1 Y-Indel , Kits and their applications. Background technique [0002] The short tandem repeat sequence specific to the human Y chromosome is Y-STR (short tandem repeat, STR), which is unique to human males and is inherited according to paternal haplotypes. It is used in forensic paternity and individual identification, human populations, class origin and evolution The research has unique application value, and it also has important application prospects in the research work of non-invasive prenatal genetic diagnosis. Y-STR test technology is a supplementary method to autosomal STR test technology. This technology has very important application value in the family investig...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6888C12Q1/6844C12N15/11
CPCC12Q1/6844C12Q1/6888C12Q2600/156C12Q2563/107
Inventor 于在亮郭丹陈初光杨凡冯晓琴肖宇清
Owner SUZHOU MICROREAD GENETICS
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