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A dna test reagent and special primers for sample screening

A reagent and sample technology, applied in the field of DNA testing reagents and special primers, can solve the problems of sample cross-contamination, increased testing costs, and increased workload of testing personnel

Active Publication Date: 2018-03-23
INST OF FORENSIC SCI OF MIN OF PUBLIC SECURITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

There are two main problems in the large-scale inspection process. One is that it is impossible to determine whether there is sample cross-contamination in a large number of samples with the same Y-STR typing; Samples undergo autosomal STR testing for precise alignment
This will greatly increase the workload of inspectors, reduce overall inspection efficiency, and increase inspection costs

Method used

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  • A dna test reagent and special primers for sample screening
  • A dna test reagent and special primers for sample screening
  • A dna test reagent and special primers for sample screening

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0071] Example 1. Preparation of DNA testing reagents for large-scale sample screening

[0072] 1. Determination of sites for DNA testing for large-scale sample screening

[0073] Since Y-STR has the characteristics of paternal inheritance, under normal circumstances, the STR typing results obtained from the samples of the same family are exactly the same. There are two main problems in the large-scale inspection process. One is that it is impossible to determine whether there is sample cross-contamination in a large number of samples with the same Y-STR typing; Samples were subjected to autosomal STR testing for accurate alignment.

[0074] Therefore, 23 loci were determined for DNA testing for large-scale sample screening as follows: 21 Y-STR loci, 1 sex locus Amelogenin, and 1 autosomal STR locus D18S51.

[0075] 上述21个Y-STR位点分别为DYS460、DYS458、DYS389I、DYS438、DYS389II、DYS390、DYS392、DYS393、DYS437、DYS385a / b、YGATA_H4、DYS391、DYS447、DYS19、DYS448、DYS456、DYS635、DYS439、DYS527a / b; ...

Embodiment 2

[0100] Example 2. Application of DNA testing reagents for large-scale sample screening

[0101] 1. Sample preparation

[0102] The samples are accumulated from the daily DNA database construction of the Physical Evidence Identification Center of the Ministry of Public Security. The samples are all paper type, blood collection cards.

[0103] Use a 0.5mm hand-held puncher to punch out a disc (no more than 1.2mm) from the blood collection card (known to be male), and place it in a 0.2mL thin-walled tube.

[0104] 2. Sample amplification

[0105] Add 10 ul of the DNA test reagent prepared in Example 1 into the above-mentioned thin-walled tube with the blood collection card for PCR amplification.

[0106] The PCR amplification program was: 95°C, 11min; 94°C, 30s, 59°C, 2min, 72°C, 1min, 28 cycles; 60°C, 1h, 25°C, forever.

[0107] 3. PCR product detection

[0108] Take 1 μL of the above-mentioned amplification product and add 10 μL Hi-Di formamide (ABI Company), and place it ...

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Abstract

The invention discloses a DNA testing reagent and special primers for sample screening. The present invention provides a DNA test primer set, consisting of a primer pair set for amplifying the following 21 Y-STR sites, a primer pair for amplifying sex locus Amelogenin, and a primer for amplifying autosomal STR sites pair composition. Experiments of the present invention prove that the DNA detection reagent provided by the present invention can simultaneously check 21 Y-STR loci and 1 autosomal STR locus, and the primers have high specificity, can obtain complete STR typing, and peak type Sharp and clear, well-balanced, no Pull-up peaks, stutter bands, and non-specific artifacts, it can fully meet the requirements of large-scale forensic DNA sample testing.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a DNA testing reagent and special primers for sample screening. Background technique [0002] Since the advent of DNA testing technology, especially (Short Tandem Repeats) STR testing technology in the mid-1980s, it has been used in criminal and civil cases, large-scale catastrophic accidents (such as: the US 9.11 terrorist attack, Indonesian tsunami, Wenchuan Earthquake) and It has played a huge role in mass emergencies. Short tandem repeat (STR), also known as microsatellite DNA (microsatellite DNA), STR detection technology mainly detects the polymorphism of these specific STR sites, after obtaining the information of multiple corresponding STR sites , to perform parallel comparison of multi-locus information, thereby realizing individual identification and paternity identification. With its unique technical advantages, STR detection has been the focus and core of forensic genet...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6888C12N15/11
Inventor 赵兴春叶健姜伯玮欧元
Owner INST OF FORENSIC SCI OF MIN OF PUBLIC SECURITY
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