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Fluorescently-labeled insertion/deletion (InDel) genetic polymorphism locus composite amplification system and application thereof

A compound amplification system and the technology of the compound amplification system are applied in the field of biological detection to achieve the effects of high polymorphism, accurate typing results and good repeatability

Active Publication Date: 2012-07-25
ACADEMY OF FORENSIC SCIENCE
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Problems solved by technology

At present, a fluorescent-labeled indel genetic polymorphism marker multiple amplification system developed for the Chinese population has not been reported

Method used

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  • Fluorescently-labeled insertion/deletion (InDel) genetic polymorphism locus composite amplification system and application thereof

Examples

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Effect test

Embodiment 1

[0051] Example 1 Application of InDel Site Fluorescence Multiplex Amplification System for Paternity Test and Individual Identification

[0052] Steps:

[0053] 1. DNA extraction

[0054] Genomic DNA extraction by Chelex-100 method: add 1mL double-distilled water to a 1.5mL centrifuge tube, then add 3μL whole blood or 3×3mm blood spot, mix carefully; incubate at room temperature for 30min, shake intermittently; centrifuge at 14,000 r / min for 5min; Carefully remove the supernatant. Add 200 μL of 10% Chelex 100 (100mesh) to the pellet; incubate at 56°C for 30 min; shake at high speed for 5-10 sec; bath in boiling water for 8 min; centrifuge at 14,000 r / min for 5 min, and take the supernatant for PCR amplification.

[0055] 2. PCR amplification

[0056] The total reaction volume was 15.0 μL, including 7.5 μL of PCR reaction buffer (Buffer), 3.0 μL of primer mix (Primer pair Mix), 0.5 μL of DNA polymerase, 3.0 μL of template DNA, and 1 μL of ultrapure water.

[0057] In the...

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Abstract

The invention belongs to the technical field of biological detection, and in particular relates to a fluorescently-labeled insertion / deletion (InDel) genetic polymorphism locus composite amplification system and application thereof. The application comprises the following steps of: selecting 35 InDel loci, designing polymerase chain reaction (PCR) amplification primers of the 35 InDel loci, matching fluorescein labels of the PCR primers, compositely amplifying and detecting the 35 InDel loci, and the like. The system can analyze 35 InDel genetic polymorphism loci by the composite amplification. The 35 InDel loci are labeled by FAM, HEX, TAMRA and ROX fluorescein with four different colors respectively. The composite amplification system based on the invention can be made into a kit, serves as an InDel locus fluorescence composite amplification kit with Chinese characteristics, and is applied to the fields, such as triplet paternity test, doublet paternity test, grandparent and grandchild test, sibling test, individual recognition, disease diagnosis, anthropology and the like.

Description

technical field [0001] The invention belongs to the technical field of biological detection, and relates to the detection of polymorphic genetic markers in the human genome, in particular to a fluorescence detection method carried out by compound amplification of 35 insertion-deletion genetic polymorphic markers (Insertion / Deletion, InDel). Labeled multiplex amplification system and its application. Background technique [0002] STR locus (short tandem repeat, STR) is a kind of DNA tandem repeat sequence widely present in eukaryotic genomes. Its core sequence is 2-7bp, and the number of repeats is usually 15-30 times. The high polymorphism of the STR locus and its following the Mendelian co-dominant inheritance law in the process of gene transmission make the PCR-STR composite amplification fluorescence detection technology an indispensable and important technical means in the international forensic field , is playing an increasingly important role in judicial practices suc...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/68G01N21/64
Inventor 赵书民李成涛张素华
Owner ACADEMY OF FORENSIC SCIENCE
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