Fluorescent multiplex amplification system of 54 Y chromosome loci, kit and application

A compound amplification system and Y chromosome technology, applied in the field of biological detection, can solve the problem that the locus information cannot meet the requirements of practical applications, etc., achieve high cumulative individual recognition and non-parent exclusion rate, improve utilization rate, and stability Good results

Pending Publication Date: 2021-04-16
SUZHOU MICROREAD GENETICS
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The number of loci contained in the Y chromosome STR kits currently on the market is small, and the locus information obtained by one amplification may not meet the requirements of pract...

Method used

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  • Fluorescent multiplex amplification system of 54 Y chromosome loci, kit and application
  • Fluorescent multiplex amplification system of 54 Y chromosome loci, kit and application
  • Fluorescent multiplex amplification system of 54 Y chromosome loci, kit and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0079] Example 1: Amplification of DNA, blood card and saliva card

[0080] Take 1 ng of commonly used DNA 9948, male blood card and saliva card as templates, mix each reaction reagent according to the determined ratio, oscillate and centrifuge, and put it into a PCR instrument for amplification.

[0081] Capillary electrophoresis detection after amplification: mix the internal standard QD650 and formamide at a ratio of 3:100 and centrifuge briefly, take 9ul of the mixture and add it to a 96-well plate, then add 1μl of the amplified product sample or allele standard, Mix and let stand for several minutes, denature at 95°C for 3 minutes, immediately bathe in ice for 3 minutes, centrifuge and put on ABI3500 sequencer for detection.

[0082] Import the raw data into the GeneMapper ID-X software, and use the GeneMapper ID-X software to analyze the obtained data and generate maps.

[0083] Depend on Figure 1-Figure 5 From the results shown, it can be seen that using the system o...

Embodiment 2

[0084] Embodiment 2: Sensitivity detection

[0085] Sensitivity detection was performed using a fluorescent multiplex amplification system with 54 sites, and 9948 was used as a template for amplification. The detection spectrum is as follows Figure 6 As shown, the minimum detection sensitivity of this system is 0.06ng, and all types can be completely detected at 0.06ng.

Embodiment 3

[0086] Example 3: Species-specific detection

[0087] The specific detection of different species is carried out by using the fluorescent multiplex amplification system with 54 sites. The DNA templates of mice, pigs, dogs, cattle and sheep are taken, and the detection patterns are as follows: Figure 7 As shown, this system does not amplify bands in other species, indicating better species specificity.

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Abstract

The invention belongs to the technical field of biological detection, and discloses a fluorescence multiplex amplification system of 54 Y chromosome loci, a kit and application. The fluorescence multiplex amplification system comprises a plurality of pairs of specific primers, and can simultaneously amplify 53 Y chromosome STR loci and one Y-Indel. Six-color fluorescence labeling is adopted, the 53 Y chromosomes STR and the one YIndel are subjected to one-tube amplification detection, the requirements of Y-STR DNA database construction of the Ministry of Public Security are met, 20 recommended cores, 15 preferred loci and some alternative loci are included, and high cumulative individual recognition capacity and cumulative non-parent exclusion rate are achieved; and meanwhile, the kit has the characteristics of large information amount and good compatibility, can directly amplify FTA cards, saliva cards and extracted DNA samples, and is simple and convenient to operate.

Description

technical field [0001] The invention belongs to the technical field of biological detection and is used for detecting the polymorphism of short tandem repeat sequence loci in the human genome. Fast and accurate judgment, and then provide judgment basis for forensic individual identification and paternity test. Background technique [0002] Forensic DNA testing technology is one of the widely used technical means in the field of forensic science, and it has played an important role in practical work. In the usual genomic DNA test, the test objects are divided into autosomes and sex chromosomes, where the sex chromosomes include X and Y chromosomes. DNA testing for the Y chromosome can carry out family investigation and assist paternal kinship identification, and then help limit the family group of the suspect and provide important clues for case investigation. [0003] The Y chromosome STR locus refers to a short tandem repeat sequence consisting of 2-6 nucleotides existing...

Claims

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Application Information

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IPC IPC(8): C12Q1/6888C12Q1/6858C12N15/11
Inventor 于在亮李思思郭丹高运通钱勇
Owner SUZHOU MICROREAD GENETICS
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