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A kind of str kit and its application for trace and degraded samples

A kit and micro-quantity technology, applied in the field of molecular biology, can solve the problems of destroying the PCR amplification process, failure to obtain typing results, multi-inhibitor components, etc., to increase the number of detections, rapid detection, and anti-inhibition capabilities strong effect

Active Publication Date: 2019-10-15
AGCU SCIENTECH +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Some samples have special sources such as blood stains in the soil, etc. Even after the DNA extraction step, there are still a large number of PCR inhibitors such as heme, humic acid, etc., which destroy the PCR amplification process, resulting in the inability to obtain a complete analysis. type result
Crude DNA extraction methods such as Chelex will also leave more inhibitor components

Method used

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  • A kind of str kit and its application for trace and degraded samples
  • A kind of str kit and its application for trace and degraded samples
  • A kind of str kit and its application for trace and degraded samples

Examples

Experimental program
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Effect test

Embodiment 1

[0036] Samples such as cigarette butts, bloodstains, and semen in actual cases were detected using the kit of the present invention and the kit of ID plus (ABI, an American company) respectively. The kit of the present invention consists of the following: Master Mix, 2mL; Genomic DNA, 10ng; Primer Mix, 1mL; sdH 2 0, 2mL; wherein, the Reaction Mix composition is: MgCl 2 7.5mM, pH8.3 Tris-HCl 125mM, KCl 125mM, dNTPs 7.5mM, BSA 2mg / ml, 5U / μL hot-start Taq enzyme, PCR enhancer, the enhancer can be trehalose, polyethylene with a concentration lower than 5% Diol, dimethyl sulfoxide, NP40, etc. IDplus uses ABI finished kit standard system and procedures.

[0037] 1. Samples of various inspection materials are provided by Suzhou Municipal Public Security Bureau.

[0038] 2. Genomic DNA extraction of various specimens

[0039] The extraction of genomic DNA from various specimens was carried out with reference to "GA / T 383-2002 Forensic Science DNA Laboratory Test Standards", using...

Embodiment 2

[0055] Various traces, wipes, and old degradation samples in actual cases were detected using the kit of the present invention and the kit of ID plus (ABI, an American company) respectively.

[0056] The kit of the present invention consists of the following: Master Mix, 2mL; Genomic DNA, 10ng; Primer Mix, 1mL; sdH 2 0, 2mL; wherein, the Reaction Mix composition is: MgCl 2 7.5mM, pH8.3Tris-HCl 125mM, KCl 125mM, dNTPs 7.5mM, BSA 2mg / ml, 5U / μL hot-start Taq enzyme, PCR enhancer, the enhancer can be trehalose, polyethylene glycol with a concentration lower than 5% Alcohol, dimethyl sulfoxide, NP40, etc.

[0057] 1. Samples of various inspection materials are provided by Suzhou Municipal Public Security Bureau.

[0058] 2. Genomic DNA extraction of various samples

[0059] The extraction of genomic DNA from various samples was carried out referring to "GA / T 383-2002 Forensic Science DNA Laboratory Test Specification", and all such samples were extracted by the magnetic bead me...

Embodiment 3

[0075] Mixed samples in actual cases are tested using the kit of the present invention.

[0076] The kit of the present invention consists of the following: Master Mix, 2mL; Genomic DNA, 10ng; Primer Mix, 1mL; sdH 2 0, 2mL; wherein, the Reaction Mix composition is: MgCl 2 7.5mM, pH 8.3 Tris-HCl 125mM, KCl 125mM, dNTPs 7.5mM, BSA 2mg / ml, 5U / μL hot-start Taq enzyme, PCR enhancer, the enhancer can be trehalose, polyethylene glycol with a concentration lower than 5% , dimethyl sulfoxide, NP40, etc.

[0077] 1. Samples of various inspection materials are provided by Suzhou Municipal Public Security Bureau.

[0078] 2. Genomic DNA extraction of various samples

[0079] The extraction of genomic DNA from various specimens was carried out referring to "GA / T 383-2002 Forensic Science DNA Laboratory Test Specification", and all such samples were extracted by the magnetic bead method.

[0080] 3. The operation steps are as follows:

[0081] 3.1 The amplification system is:

[0082] ...

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Abstract

The invention discloses an STR kit for a micro-amount degradation sample and application of the STR kit. The kit comprises 17 pairs of specific amplification primers of STR gene loci, the 17 pairs of specific amplification primers are divided into 5 groups, and fluorescent labels with 6 colors are related. The STR kit has the advantages of rapidness, high sensitiveness and high inhibition resistance on detection of micro-amount degradation samples. A mini type amplification system of the 17 gene loci established by the invention is short in amplified fragment, and is suitable for micro-amount, highly degraded or high-inhibitor-content difficult samples; and during actual application, STR types of special biological samples such as skeletons, contact cast-off cells and blood samples in soil can be obtained successively. DYS391 gene loci are introduced in a system, and the auxiliary judgment effect on sex genes Amelogenin can be achieved; and the kit can rapidly detect the samples, and the whole PCR process does not exceed 50 minutes.

Description

technical field [0001] The invention belongs to the field of molecular biology, relates to molecular genetics, in particular to an STR kit for trace and degraded samples and its application. 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 FBI selected 13 STR loci for the establishment of a DNA database—CODIS (Combined DNA Index System): CSF1PO, D3S1358, D5S818, D7S820, D8S1179, D13S317, D16S539, D18S...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6888
Inventor 周如华石云杰张健王鑫孙元鹏陈林丽张雷王艳芳郭育林周翔郑卫国
Owner AGCU SCIENTECH
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