Fluorescence multiplex amplification system, kit and application thereof

A compound amplification system and kit technology, applied in biochemical equipment and methods, microbial measurement/inspection, etc., can solve the problems of inability to apply DNA library construction, high price of single-tube detection, expensive imported reagents, etc., to achieve Unique and novel kit, high sample applicability, and short detection cycle

Active Publication Date: 2021-08-27
SUZHOU MICROREAD GENETICS
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] At present, chip analysis or sequencing is mostly used in domestic detection of SNP sites, which takes a long time and is expensive; SNP site detection for in vitro diagnosis mostly uses the Taqman probe method, which is easy to operate, but single-tube detection is expensive and difficult to use Forensic database
AB has a kit SNaPshot that combines SNP and forensic medicine. It performs single base extension reaction and detects it with 3730 sequencer. Also cannot be applied to DNA library construction

Method used

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  • Fluorescence multiplex amplification system, kit and application thereof
  • Fluorescence multiplex amplification system, kit and application thereof
  • Fluorescence multiplex amplification system, kit and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0080] (1) Obtain samples: (a) Genomic DNA was extracted from the following samples by Chelex-100 method or magnetic bead method, (b) whole blood, (c) blood card, (d) saliva card, (e) hair with follicle ;

[0081] (2) Shake and mix each reaction reagent and make PCR premix solution according to the determined concentration. The total volume of the amplification system is 24 μl, including 5 μl of 24 pairs of specific primer mixtures carrying fluorescent labels, and make a kit, and then add it to the kit Inject 1 μl of the collected genomic DNA.

[0082] (3) Carry out PCR amplification according to the determined reaction conditions.

[0083] (4) Capillary electrophoresis detection: Mix the QD550 internal standard and formamide in a ratio of 3:100, take 10ul of the mixture and add it to a 96-well plate, then add 1μl of the amplification product sample or allelic standard, mix and stand for a few minutes , denatured at 95°C for 3 minutes, immediately bathed in ice for 3 minutes...

Embodiment 2

[0086] Embodiment 2: Utilize the fluorescence multiplex amplification kit of 24 Y-SNP loci to carry out species-specific detection, the species detected in the present invention is monkey-derived cells, and the detection results are as follows Figure 8 As shown, no non-specific bands appeared.

Embodiment 3

[0087] Embodiment 3: Utilize the fluorescent multiplex amplification kit of 24 Y-SNP sites to carry out repeated freezing and thawing tests, and the detection pattern is as follows Figure 9 As shown, there was no change in the amplification effect after repeated freezing and thawing for ten times.

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Abstract

The invention discloses a fluorescent composite amplification system, a kit and applications thereof. The fluorescent composite amplification system includes multiple pairs of specific primers, the nucleotide sequence of which is shown in SEQ ID NO.1-71, and the The fluorescent compound amplification system is used to make a kit, and the prepared kit is applied to forensic DNA analysis and ancestry analysis. The Y-SNP provided by the present invention includes many haplogroups, has a high degree of differentiation for male groups, and the kit is unique and novel, with a large amount of information and good compatibility.

Description

technical field [0001] The invention belongs to the technical field of biological detection, and in particular relates to a fluorescent compound amplification system, a kit and an application thereof. Background technique [0002] At present, DNA detection technology has been widely used in forensic DNA analysis, genetic relationship identification, cell line identification, medical genetics, anthropology and other fields. The most widely used DNA analysis is the analysis of genetic markers, and the core technology of most DNA analysis methods lies in the analysis of fragment length polymorphism and sequence polymorphism. [0003] SNP research is an important step towards the application of the Human Genome Project. This is mainly because SNP will provide a powerful tool for the discovery of high-risk groups, the identification of disease-related genes, the design and testing of drugs, and basic research in biology. SNPs are widely distributed in the genome, and studies ha...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6858C12Q1/6888
CPCC12Q1/6858C12Q1/6888C12Q2600/156C12Q2600/16C12Q2535/137C12Q2531/113C12Q2537/143C12Q2563/107C12Q2565/125
Inventor 于在亮杨凡郑玉高运通李逸凡陈初光张蕊
Owner SUZHOU MICROREAD GENETICS
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