DNA examination reagent for sample examination, and special primer thereof

A reagent and primer pair technology, applied in the field of DNA testing reagents and special primers, can solve the problems of increasing the testing cost, reducing the overall testing efficiency, and inability to determine the sample.

Active Publication Date: 2015-09-16
INST OF FORENSIC SCI OF MIN OF PUBLIC SECURITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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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 sa...

Method used

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  • DNA examination reagent for sample examination, and special primer thereof
  • DNA examination reagent for sample examination, and special primer thereof
  • DNA examination reagent for sample examination, and special primer thereof

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 examination reagent for sample examination, and a special primer thereof. A DNA examination primer group provided by the invention is composed of a primer pair group used for amplifying 21 Y-STR sites, a primer pair for amplifying a sex site Amelogenin, and a primer pair for amplifying an autosome SRT site. Experiments prove that the DNA examination reagent can simultaneously examine the 21 Y-STR sites and the autosome SRT site, and the primer specificity is high, so complete SRT typing can be obtained, peak patterns are sharp and intelligible, the balance is good, no Pull-up peaks or stutter bands appear, no non-specific artificial products appear, and legal medical experts' DNA large scale sample examination requirements are completely met.

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] DNA testing technology, especially the (Short Tandem Repeats) STR testing technology, has been used in criminal and civil cases, large-scale catastrophic accidents (such as: 9.11 terrorist attacks in the United States, 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 genetics research for deca...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N15/11
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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