Fluorescence labeled multiplex amplification kit for simultaneously analyzing 24 loci of human genome DNA, and application thereof

A technology of fluorescent labeling and compound amplification, which is used in the determination/inspection of microorganisms, biochemical equipment and methods, etc., and can solve the problems of one child with multiple parents in the database, and the identification of close relatives with mutations in parent-child identification.

Active Publication Date: 2016-03-09
AGCU SCIENTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The system can be applied to the public security department's daily case investigation, individual identification, paternity identification, and archaeology, etc. It helps to solve the proble

Method used

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  • Fluorescence labeled multiplex amplification kit for simultaneously analyzing 24 loci of human genome DNA, and application thereof
  • Fluorescence labeled multiplex amplification kit for simultaneously analyzing 24 loci of human genome DNA, and application thereof
  • Fluorescence labeled multiplex amplification kit for simultaneously analyzing 24 loci of human genome DNA, and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0078] Example 1 A kit for simultaneous analysis of fluorescently labeled multiplexed amplification of 24 loci of human genomic DNA, consisting of the following:

[0079] components

volume

Reaction Mix

10.0 μL

Genomic DNA

Xμl content is 0.125-5ng

Primers corresponding to the above 24 loci

5.0 μL

Hot start Taq enzyme (5U / μL)

0.5μL

wxya 2 o

Make up to 25.0 μL

[0080] The ReactionMix mentioned in it is MgCl 2 7.5mM, Tris-HClbuffer125mM, KCl125mM, dNTPs7.5mM, BSA2mg / mL.

[0081] The primers and primer concentrations corresponding to the 24 loci are:

[0082]

[0083]

[0084] The 5' end of at least one of the primers corresponding to each locus is labeled with a fluorescent dye.

[0085] The fluorescent dye marker is 6-FAM, HEX, TAMRA or ROX.

[0086] The 24 loci are grouped, and the following is one of the product combination methods: D16S539, D6S477, D15S659, D10S1248, D1S1656 and D21S2055 are...

Embodiment 2

[0100] Application of 24-locus fluorescence-labeled multiple amplification test system for triplet paternity identification

[0101] 1. Collect blood samples in paternity testing cases: the samples in this experiment are provided by XX paternity testing agency. For DNA extraction, use the Chelex-100 method to extract genomic DNA from three whole blood samples: take 0.5-5 μl of whole blood and place it in a sterilized 1.5ml centrifuge tube, add 1ml of sdH2O to the tube, shake for a few seconds; place at room temperature for 10 minutes , shake for a few seconds, centrifuge at 12,000 rpm for 3 minutes, discard the supernatant, keep enough supernatant to cover the precipitate, do not stir up the precipitate; add 200 μl of 5% Chelex-100, shake for a few seconds; incubate at 56°C for 30 minutes, shake for a few seconds; Bath in boiling water for 10 minutes, shake for a few seconds; centrifuge at 12,000rpm for 5 minutes, and the supernatant is the extracted genomic DNA. Genomic DNA ...

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Abstract

The invention discloses a fluorescence labeled multiplex amplification kit for simultaneously analyzing 24 loci of human genome DNA, and an application thereof. The 24 loci comprise D16S539, D6S477, D15S659, D10S1248, D1S1656, D21S2055, D4S2366, D2S441, D3S1744, D22-GATA198B05, D3S3045, D11S2368, D17S1290, D10S2325, D14S608, D8S1132, D7S3048, D5S2500, D7S1517, D18S535, D10S1435, D13S325, D19S253 and a sex locus Amelogenin. The 24 loci are divided into 4 groups, and fluorescence labels with 5 colors are involved; the above fluorescence labeled multiplex amplification system has high sensitivity, and can detect all the 24 loci when the DNA template amount is 0.12ng; and the fluorescence labeled multiplex amplification system is suitable for being used in blood filter paper and direct amplification of FTA card acquired samples.

Description

technical field [0001] The invention relates to a 24-locus fluorescent label composite amplification test system, in particular to a fluorescent label composite amplification kit for simultaneously analyzing 24 loci of human genome DNA and its application. Background technique [0002] Short tandem repeat loci (STR) are currently commonly used genetic markers. Compared with other genetic markers, STR markers have small fragments and are easy to amplify, and are more suitable for trace and degraded samples. The amplification conditions of each locus are similar and can be multiplexed, so it is fast, efficient, and accurate. , sensitivity, large amount of information and so on. Especially in the establishment of DNA database, compared with the previous RFLP and other technologies, STR multiplex amplification technology has great advantages. Therefore, the US FBI has conducted a lot of research and selected 13 STR loci for the establishment of a DNA database - CODIS (Combined...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
CPCC12Q1/6876C12Q2600/156
Inventor 郑卫国张雷王艳芳李科杰齐丽媛王於建郭育林
Owner AGCU SCIENTECH
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