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Mini-STR kit for trace and degradable test materials

A mini-str and kit technology, applied in the field of forensic DNA detection, can solve the problems of low detection rate and high inefficiency, achieve high sensitivity, strong adaptability, and increase the number of detections

Pending Publication Date: 2020-06-12
广州万维泰生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] The technical problem to be solved by the present invention is the low detection rate and high inefficiency of the existing DNA detection kit due to the degradation of the test material

Method used

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  • Mini-STR kit for trace and degradable test materials
  • Mini-STR kit for trace and degradable test materials

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0056] Bloodstains and difficult inspection materials (teeth, white bones, nails, exfoliated cells, etc.) samples in actual cases use the detection kit of the present invention respectively. The kit of the present invention is composed of the following: 2X PCR Mastermix, genomic DNA 100pg / uL, 5X primer mix, ddH 2 0; Wherein, described 2X PCR Master mix composition is: Mg 2+ ,Tris-HCl,K +, dNTPs, hot-start Taq enzyme, PCR enhancer, etc., the enhancer can be one, two or more of trehalose, polyethylene glycol, NP4, betaine, etc., Mg 2+ by MgC1 2 get, K + Obtained via KCl.

[0057] 1. The samples of various inspection materials were provided by the Criminal Investigation Bureau of the Guangdong Provincial Public Security Bureau, the Department of Forensic Medicine of Sun Yat-sen University, and the laboratories of the Law School of Southern Medical University.

[0058] 2. Genomic DNA extraction of various specimens

[0059] The extraction of genomic DNA from various specimen...

Embodiment 2

[0073] The actual case—the samples of obsolete inspection materials in 2001 were tested using the kit of the present invention and the kit of PP21 (Promega, USA) respectively.

[0074] The test kit of the present invention is composed of the following: 2X PCRMastermix, genomic DNA 100pg / uL, 5X primer mixture, ddH 2 0; Wherein, described 2X PCR Master mix composition is: Mg 2+ ,Tris-HCl,K + , dNTPs, hot-start Taq enzyme, PCR enhancer, etc., the enhancer can be one, two or more of trehalose, polyethylene glycol, NP4, betaine, etc., Mg 2+ by MgC1 2 get, K + Obtained via KCl.

[0075] PP21 uses its kit system and procedures as follows:

[0076] 1. The samples for inspection are provided by the Laboratory of the Criminal Investigation Bureau of the Guangdong Provincial Public Security Bureau.

[0077] 2. Genomic DNA extraction from samples

[0078] The extraction of genomic DNA from the sample was carried out according to "GAT383-2002 Forensic Science DNA Laboratory Test Spe...

Embodiment 3

[0096] Sensitivity and balance of the present invention are tested, such as Figures 15-20 shown. Use the positive DNA control 2800M and 9947A as the detection samples to carry out the sensitivity and balance detection of the kit of the present invention.

[0097] The test kit of the present invention is composed of the following: 2X PCRMastermix, genomic DNA 100pg / uL, 5X primer mix, ddH 2 0; Wherein, described 2X PCR Master mix composition is: Mg 2+ ,Tris-HCl,K + , dNTPs, hot-start Taq enzyme, PCR enhancer, etc., the enhancer can be one, two or more of trehalose, polyethylene glycol, NP4, betaine, etc., Mg 2+ Obtained by MgC12, K + Obtained via KCl.

[0098] 1. The positive DNA control samples of 2800M (10ng / uL) and 9947A (0.125ng / uL) were provided by the Criminal Investigation Bureau Laboratory of Guangdong Provincial Public Security Bureau.

[0099] 2. Concentration gradient dilution of 2800M positive DNA control sample of the test material.

[0100] Dilute 2800M 10n...

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Abstract

The invention discloses a mini-STR kit for a trace and degradable test material. The kit comprises specific amplification primers of 20 pairs of core loci, the core PCR products of the specific amplification primers are all less than 250bp; wherein the STR locus is shown in the specification; D16S539, D18S51, D21S11, D2S441, D13S317, FGA, D7S820, Amelogenin, D3S1358, D12S391, CSF1PO, D1S1656, THO1, D19S433, D8S1179, D10S1248, THOX, vWA, D5S818, and D2S1338 are used in the method. According to the invention, a mini amplification system with 20 gene loci is established, amplicon fragments are further shortened and are controlled within 250bp, and the detection number of the gene loci in a degradation detection material is increased; aiming at detection of difficult detection materials suchas trace detection materials and degradation detection materials, the method has the advantages of rapidness, high sensitivity and strong adaptability; the method is suitable for difficult samples which are highly degraded or have high inhibitor content, and can successfully obtain STR typing of special biological samples such as avicennia corpses, teeth, toenails, contact cast-off cells, old casesamples and the like in practical application.

Description

technical field [0001] The invention relates to the field of forensic DNA detection, in particular to a mini-STR kit for trace and degraded test materials. Background technique [0002] The database construction of STR DNA is gradually standardized in my country. According to the latest public security industry standard of the People's Republic of China, there are 20 autosomal core loci in China. In recent years, the US FBI has also increased the number of core loci to 20 on the basis of the original 13 CODIS loci. In addition, Europe has also promulgated European standards ( ESS), the union of these core loci is Amelogenin, D18S51, D21S11, D3S1358, FGA, D8S1179, vWA, CSF1PO, D16S539, D7S820, D13S317, D5S818, D2S1338, D19S433, TH01, TPOX, D6S1043, Pent D, Penta E, D12S391, D1S1656, D2S441, D22S1045, D10S1248 and SE33. Therefore, the kits used by major manufacturers for the construction of DNA databases basically cover the above-mentioned loci, such as ABI AmpFeSTRglobal fi...

Claims

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Application Information

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IPC IPC(8): C12Q1/6888
CPCC12Q1/6888C12Q2600/156
Inventor 刘建世赵凯
Owner 广州万维泰生物科技有限公司
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