A kit for simultaneous analysis of 26 loci of human genome DNA by fluorescent labeling compound amplification and its use method and application

A technique for fluorescent labeling and compound amplification, applied in the field of five-color fluorescent label compound amplification system

Active Publication Date: 2016-04-20
AGCU SCIENTECH +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

This has a certain impact on the application and efficiency of DNA testing technology

Method used

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  • A kit for simultaneous analysis of 26 loci of human genome DNA by fluorescent labeling compound amplification and its use method and application
  • A kit for simultaneous analysis of 26 loci of human genome DNA by fluorescent labeling compound amplification and its use method and application
  • A kit for simultaneous analysis of 26 loci of human genome DNA by fluorescent labeling compound amplification and its use method and application

Examples

Experimental program
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Effect test

Embodiment 1

[0066] Example 1 A kit for simultaneous analysis of fluorescently labeled multiplexed amplification of 26 loci of human genomic DNA, consisting of the following:

[0067] components

volume

Reaction Mix

10.0 μL

Primers corresponding to 26 loci

5.0 μL

Hot start Taq enzyme (5U / μL)

0.5μL

wxya 2 o

Make up to 25.0 μL

[0068] The ReactionMix mentioned in it is MgCl2 7.5mM, Tris-HClbuffer125mM, KCl125mM, dNTPs7.5mM, BSA2mg / mL.

[0069] The primers and primer concentrations corresponding to the 26 loci are:

[0070]

[0071] .

[0072] The 5' end of at least one of the primers corresponding to each locus is labeled with a fluorescent dye.

[0073] The fluorescent dye marker is 6-FAM, HEX, TAMRA or ROX.

[0074] The 26 loci are grouped, specifically: D3S1358, D13S317, D7S820, D16S539, D1S1656, and PentaE are the first group, and the fluorescent markers of the primers corresponding to this group of loci are those of ...

Embodiment 2

[0089] Application of 26-loci fluorescence-labeled multiple amplification test system for triplet paternity identification

[0090] 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 12,000rpm for 3 minutes, discard the supernatant, keep enough supernatant to cover the precipitate, do not stir 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 was e...

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Abstract

The invention relates to a five-color fluorescent mark composite amplification system for simultaneous detection of 26 loca of human genome DNA. The 26 loca are divided into four groups, and fluorescent marks in five colors are involved totally. The five-color fluorescent mark composite amplification system is high in sensitivity; under the condition that the DNA template amount is 0.12ng, all the 26 loca can be detected; the five-color fluorescent mark composite amplification system is suitable for direct amplification of blood filter paper and FTA card collection samples.

Description

technical field [0001] The present invention relates to a 26-loci fluorescent marker composite amplification inspection system, in particular to a five-color fluorescent marker composite amplification system for simultaneously analyzing 26 loci of human genome DNA, which can be applied to individual identification and paternity identification . Background technique [0002] Short tandem repeat loci (STRs) are currently commonly used genetic markers. In the early 1990s, the polymorphism of the STR locus was discovered, especially the STR locus has a small fragment and is easy to amplify, which is suitable for testing trace and degraded samples, and the amplification conditions of each locus are similar and can be compounded and amplified, so it has the advantages of Sensitive, accurate, fast, large amount of information and other advantages. Especially in the establishment of DNA database, STR compound amplification technology has great advantages. Therefore, the US FBI ha...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/68
Inventor 郑卫国卢文翔杜蔚安卢青葛海鹏薛佳郭育林朱丽君许峻荣
Owner AGCU SCIENTECH
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