ZNA (zip nucleic acid) primer based STR (short tandem repeat) typing kit

A kit and forward primer technology, which is applied in the field of forensic DNA testing and analysis, can solve problems such as poor typing effect, inability to meet testing requirements at the same time, and unsuitability for the Chinese population.

Active Publication Date: 2016-11-09
BEIJING INST OF GENOMICS CHINESE ACAD OF SCI CHINA NAT CENT FOR BIOINFORMATION
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  • Abstract
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AI Technical Summary

Problems solved by technology

This strategy also has some shortcomings in practical application: allele loss occurs; the whole genome amplification technology for specific types of samples is not yet clear, etc.
Although it contains a relatively large number of STR loci, its applicability to different populations in China is weak; and when the template amount is less than 0.1ng, the typing effect is not good
[0008] In summary, although STR typing technology is of great significance in the field of forensic DNA testing and analysis, most of the currently established commercial amplification systems or kits are aimed at Western populations, and are not suitable for Chinese populations in actual use; A small number of detection systems suitable for the Chinese population, on the one hand, cannot meet the detection of trace (or low copy number) and highly degraded biological samples at the same time, on the other hand, the general applicability to various groups of people is poor

Method used

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  • ZNA (zip nucleic acid) primer based STR (short tandem repeat) typing kit
  • ZNA (zip nucleic acid) primer based STR (short tandem repeat) typing kit
  • ZNA (zip nucleic acid) primer based STR (short tandem repeat) typing kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0153] Example 1 Genotyping of 10 different volunteers

[0154] 1. Sample collection: The sample is blood from volunteers, 10 samples in total.

[0155] 2. Template DNA extraction: Template DNA was extracted from 10 different volunteers' blood samples by Chelex-100 method.

[0156] 3. PCR amplification:

[0157] (1), the ZNA primers used are:

[0158] A pair of ZNA primers used to amplify D20S1082 are: Forward primer: The nucleic acid sequence shown in SEQ ID NO: 1 is a ZNA sequence composed of a Z unit coupled to the 9th and 15th nucleotides from the 5' end, Reverse primer: a ZNA sequence composed of a Z unit coupled to the 5th, 11th and 15th nucleotides from the 5' end of the nucleic acid sequence shown in SEQ ID NO: 2;

[0159] A pair of ZNA primers used to amplify D6S474 are: Forward primer: The nucleic acid sequence shown in SEQ ID NO: 3 is a ZNA sequence composed of a Z unit coupled to the 5th and 21st nucleotides from the 5' end, Reverse primer: the nucleic acid seq...

experiment example 2

[0186] Experimental example 2 template concentration

[0187] Taking the D20S1082 locus as an example, the blood DNA of a volunteer in Example 1 was used as a template, and its ZNA primer and DNA primer were used respectively, and the template amounts were 200, 100, 50, 25, 12.5, and 6.25 pg respectively. PCR amplification, PCR amplification system and amplification program are with embodiment 1, amplified product carries out agarose gel electrophoresis, the result is as follows figure 2 shown. Under the same amplification conditions, the concentration of the amplification product obtained by ZNA primers was significantly higher than that of DNA primers, and ZNA primers could still see clear amplification bands when the template amount was 12.5pg, while DNA primers, under the same conditions, The amplified bands with a template amount of 50 pg began to be blurred. The template concentration of the remaining loci was the same as that of the D20S1082 locus.

[0188] case:

...

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Abstract

The invention belongs to the field of forensic DNA examination and analysis and in particular relates to a miniSTR (short tandem repeat) typing kit used for trace degradation of biological samples. The kit adopts ZNA primers instead of traditional DNA primers and simultaneously carries out multiplex amplification on 12 miniSTR loci and a gender recognition locus, wherein the 12 miniSTR loci are D20S1082, D6S474, D12ATA63, D9S1122, D2S1776, D1S1627, D3S4529, D2S441, D4S2364, D3S3053, D17S1301 and D21S11; and the gender recognition locus is Amelogenin. The STR typing kit provided by the invention combines the miniSTR typing technology with ZNA primer amplification, is suitable for the Chinese population and the minority population, is suitable for trace and high degradation of biological samples and has the effect of greatly improving the typing sensitivity and accuracy.

Description

technical field [0001] The invention belongs to the field of forensic DNA testing and analysis, and in particular relates to a miniSTR typing kit for microdegrading biological samples. Background technique [0002] Short tandem repeat (short tandem repeat, STR), also known as microsatellite DNA (microsatellite DNA), is a type of genetic marker widely distributed in the human genome, and its core sequence generally consists of 2 to 6 bases. Different numbers of repeat units in the sequence result in the presence of different alleles at the same locus. Because of its relatively simple repeating unit, it is also called simple sequence repeats (SSRs). Due to the change of the number of core units, the length of STR varies greatly in the population, which constitutes the genetic polymorphism of STR, and then becomes the biological basis of STR analysis technology. The vast majority of STR sequences are distributed in the non-coding regions of the genome, the core repeat sequenc...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
CPCC12Q1/6888C12Q2600/156
Inventor 严江伟杨雅冉杨猛赵晶陈彤
Owner BEIJING INST OF GENOMICS CHINESE ACAD OF SCI CHINA NAT CENT FOR BIOINFORMATION
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