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Fluorescence labeling composite amplification detection system with 18 loci

A compound amplification system and compound amplification technology, applied in the field of fluorescent label compound amplification inspection system

Inactive Publication Date: 2009-05-27
AGCU SCIENTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

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

Method used

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  • Fluorescence labeling composite amplification detection system with 18 loci
  • Fluorescence labeling composite amplification detection system with 18 loci
  • Fluorescence labeling composite amplification detection system with 18 loci

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] Example 1 The multiplex amplification and fluorescence detection of the fluorescently labeled multiplexed amplification test system for 18 loci.

[0042] 1. PCR amplification system

[0043]

[0044] 2. Amplification thermal cycle

[0045] (1) Place the PCR amplification tube on the thermal cycler;

[0046] (2) Select the program recommended below to amplify;

[0047] (3) The amplified samples should be kept away from light;

[0048] Amplification program for thermal cycler

[0049]

[0050] 3. Fluorescent detection of the amplified product on a genetic analyzer

[0051] The sample loading mixture is composed of deionized formamide and molecular weight internal standard (AGCU Marker SIZ-500) in the system [(0.5μl AGCU Marker SIZ-500)×(injection number)+(12μl deionized formamide)×(injection number )]. Mix 12.5 μl of the loading mixture with 1 μl of the amplification product or allelic analysis standard (17+1 Allelic Ladder) in the system, avoiding the generat...

Embodiment 2

[0055] Application of 18-loci fluorescence-labeled multiple amplification test system for paternity identification

[0056] 1. Collection of blood samples in paternity test cases: The samples in this experiment were provided by the Shenzhen Blood Bank.

[0057] 2. Chelex-100 method to extract genomic DNA

[0058] Take 0.5~5μl whole blood or 1~3mm×2~5mm blood spot into a sterilized 1.5ml centrifuge tube, add sdH 2 O 1ml in 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 sediment, do not stir the sediment. Add 200 μl of 5% Chelex-100 and shake for a few seconds. Incubate at 56°C for 30 minutes, shake for a few seconds. Boiling water bath for 10 minutes, shake for a few seconds. Centrifuge at 12,000 rpm for 5 minutes, and the extracted human genomic DNA is in the supernatant.

[0059] 3. Amplification detection

[006...

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Abstract

The invention relates to a five-color fluorescence-labeling composite amplification system simultaneously analyzing 18 loci of human genomic DNA. The system divides the 18 loci into four groups, and relates to fluorescence labels in five colors in all. The fluorescence-labeling composite amplification system has the advantages of high sensitivity and capability of detecting the whole 18 loci under the condition that DNA template quantity is 0.2 ng. In Chinese Han population, the overall random matching probability is 8.8*10<-21>, and the cumulative non-parentage exclusion rate is 0.999999999993.

Description

technical field [0001] The present invention relates to a fluorescent marker composite amplification inspection system with 18 loci, in particular to a five-color fluorescent marker composite amplification system for simultaneously analyzing 18 loci of human genome DNA, which can be applied to individual identification and paternity identification . Background technique [0002] Short tandem repeat loci (STR) 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 FB...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68G01N21/64
Inventor 陈光辉熊勇华夏子芳杜蔚安郑卫国
Owner AGCU SCIENTECH
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