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Reagent and kit for detecting 13 CODIS STR gene loci of trace mixed human source DNA sample and application thereof

A locus and kit technology, which is applied in the field of amplification and detection of human short tandem repeat sequences, can solve the problems of inability to distinguish genotypes, high requirements for detection of DNA sample abundance, etc., and achieve the effect of improving the detection limit.

Inactive Publication Date: 2018-01-09
SUZHOU GENO TRUTH BIOTECHNOLOGY CO LTD +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

NGS can increase the amount of information to a certain extent, making the individual identification of the detection system stronger, but NGS has high requirements for the abundance of DNA samples, and cannot distinguish the genotyping of mixed samples

Method used

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  • Reagent and kit for detecting 13 CODIS STR gene loci of trace mixed human source DNA sample and application thereof
  • Reagent and kit for detecting 13 CODIS STR gene loci of trace mixed human source DNA sample and application thereof
  • Reagent and kit for detecting 13 CODIS STR gene loci of trace mixed human source DNA sample and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0091] 1. Sample collection:

[0092] Mixed exfoliated cell wipes from the scene of the case collected by the Public Security Bureau

[0093] 2. DNA extraction:

[0094] Use magnetic bead method to extract exfoliated cell wipes

[0095] 3. PCR system construction:

[0096] After vortexing and mixing each reaction component, it is collected by centrifugation briefly, and it is configured into PCR reaction mixture. Aliquot PCR reaction mixture, dispense 19μL into PCR reaction tube, and finally add 1μL DNA solution to the reaction system. After vortexing and mixing, centrifuge to collect, make droplets, and put Barcord label into PCR machine for reaction.

[0097] Component name

Add volume

2 ⅹ PCR reaction buffer

10μL

5ⅹ Primer combination

2μL

Taq DNA polymerase

0.2μL

DNA template

1μL

Ultra-pure water

6.8μL

[0098] 4. Thermal cycle parameters

[0099] Step 1: Denaturation at 98°C for 30 seconds, Step 2: Denaturation at 98°C for 10 seconds, Step 3: Annealing at 60°C for...

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Abstract

The invention discloses a reagent and a kit for detecting 13 CODIS STR gene loci of a trace mixed human source DNA sample and an application thereof. The reagent comprises amplification primers for 13CODIS STR gene loci and a sex gene locus, wherein the 13 CODIS STR gene loci are separately TPOX, DS31358, FGA, D5S818, CSF1PO, D8S1179, D7S820, TH01, vWA, D13S17, D16S539, D18S51 and D21S11, and thesex gene locus is Amelogenin. The sequences of the amplification primers are separately shown as SEQ ID NO: 1-SEQ ID NO: 28. The 13 CODIS STR gene loci are independent of one another without a chaineffect, and can be matched with the gene loci of an existing commercial kit and a database of the Ministry of Public Security, so that the adaptability of an analytical result is achieved; meanwhile,a single DNA molecule can be amplified and detected; on the premise of ensuring the efficiency of the system, the detection rate of degraded / broken DNA templates can be increased; the reagent and kitcan be widely applied to the fields of individual recognition by legal medical experts, paternity test and other group genetics analysis.

Description

Technical field [0001] The invention relates to the amplification and detection of human short tandem repeats (STR), in particular to a reagent, kit and application for detecting 13 CODIS STR loci in a trace mixed human DNA sample, and belongs to the field of biotechnology. Background technique [0002] Since the application of DNA atlas in forensic DNA analysis technology in 1985, DNA analysis technology has been widely used in forensic material evidence identification and has provided important evidence in many major criminal cases and civil disputes. At present, the multi-fluorescence-labeled composite amplification system based on capillary electrophoresis technology is the most widely used technology. [0003] Short tandem repeats (STRs) are microsatellites and are widely distributed in the human genome. STR sites have high genetic diversity in the population, so they are often used as genetic markers. STR fragments are generally less than 500 bp and are Del’s law is codomina...

Claims

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Application Information

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IPC IPC(8): C12Q1/6888
Inventor 赵洪玉张惠丹节淳皓戴敬
Owner SUZHOU GENO TRUTH BIOTECHNOLOGY CO LTD
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