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Composite amplification kit for 25 human chromosomal loci and its application

A composite amplification and locus technology, applied in the field of composite amplification kits, can solve problems such as poor compatibility of kits and limited number of STR loci, and achieve strong test material adaptability, good analysis data compatibility, and high compatibility. sexual effect

Active Publication Date: 2022-08-09
BEIJING PEOPLESPOT TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0009] In order to solve technical problems such as the limitation of the number of STR loci detected simultaneously by multiple amplification and the poor compatibility of the kits in the prior art, the present invention provides a multiple amplification kit capable of simultaneously detecting 25 chromosomal loci and its application

Method used

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  • Composite amplification kit for 25 human chromosomal loci and its application
  • Composite amplification kit for 25 human chromosomal loci and its application
  • Composite amplification kit for 25 human chromosomal loci and its application

Examples

Experimental program
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Effect test

Embodiment 1

[0072] Example 1 25 chromosomal gene seat compound amplification kit development

[0073] 1. Compound amplifier design of 25 chromosomal genes

[0074] For the composite amplification system, especially the compound amplification system of 25 gene seats in the present invention, the requirements of the specificity of the primer sequence, the secondary structure, the stability of the target sequence, and the efficiency of the amplification efficiency are extremely high. Continuous amplification -optimization -amplification cycle experiments have obtained 25 pairs of primers of 25 gene seats in 25 gene seats. The specific sequence list 1 is shown:

[0075] Table 1 25 genetic seats amplification primers

[0076]

[0077]

[0078] The above -mentioned primers are based on the following grouping of the corresponding gene seat: the first group is D3S1358, CSF1PO, D2S441, D21S11, Penta E; the second group is D8S1179, D5S818, D19S433, D16S539, and Penta D; No. 1 The three groups are A...

Embodiment 2

[0086] Establishment of 2 25 chromosomal gene seat compound amplification systems and procedures

[0087] Optimize the annealing temperature and cycle of the amplification primers of 25 gene seats, get the optimal response procedure of the compound amplification amplification of chromosomes: 95 ° C for 2 minutes; 94 ° C for 5 seconds, 60 ° C insulation at 60 ° C insulation In 45 seconds, 45 seconds of thermal insulation at 72 ℃, this step runs 30 cycles; 60 ° C for 20 minutes; 4-10 ° C insulation.

Embodiment 3

[0088] Example 3 Application of a composite amplification kit to generate 9948 cell strain

[0089] The kit provided by the present invention is compounded to the 9948 cell strain for compound expansion of 25 chromosomal gene seats. The CHELEX-100 method is used to extract the template DNA derived from the 9948 cell strain. The amplification reaction is performed on the ABI 9700 thermal circulation instrument. Electricity and detection are performed on the ABI 3130 genetic analyzer. The data analysis uses the Genemapper IDX V1.2 software. Reagent materials such as the genetic ladder (LADDER) used can be obtained from commercial channels and are commonly used by technicians in the field.

[0090] 1. Use the Chelex-100 method to extract DNA (refer to "Forensic DNA Protocol", Humanpress, 1998):

[0091] (1) 9948 cells cultivated by 3 μL in 500 μL of centrifugal tube.

[0092] (2) Mixed oscillation of Chelex solution so that Chelex is fully suspended, add 195 μL 5 % Chelex-100 (L00-20...

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Abstract

The present invention relates to a composite amplification kit of 25 human chromosomal loci and its application. The present invention provides a compound amplification system of 25 loci of human chromosomes, comprising specific primers for amplifying 25 loci, the 25 loci including 23 autosomal STR loci, 1 gender Identify loci and 1 Y chromosome indel site. Using six-color fluorescent labeling technology to group 25 pairs of specific primers for fluorescent labeling, through the design and optimization of primer sequences and working primer concentrations, the simultaneous efficient, specific and sensitive amplification of 25 human chromosomal loci has been achieved. The detection results of the composite amplification system have high individual identification ability and good data compatibility, and can be used for paternity testing and individual identification in practice, effectively reducing the detection cost of human DNA typing and improving the efficiency of human DNA typing. detection efficiency.

Description

Technical field [0001] The present invention involves the field of molecular genetics and biological testing technology, and specifically involves the composite amplification kit and its application of 25 personal chromosomal genes. Background technique [0002] Short series repetitive sequence (STR) is also known as microbriet or simple sequence repetition (SSR). It is a type of DNA series repetitive sequence widely existed in the nuclear genome. The core sequence is 2-6 base duplicate units. The number of STR genes is large, with a wide range of divisions, accounting for about 3 % of the entire human genome, and it is highly polymorphic. Follow Mendel's genetic laws. Therefore, STR amplification detection technology is widely used in individual identification, kinship identification, and group genetic research. [0003] At present, STR composite amplification technology is the main technical means of individual forensic identification and parent -child identification, and has b...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6888C12N15/11
CPCY02A50/30
Inventor 不公告发明人
Owner BEIJING PEOPLESPOT TECH
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