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Fluorescence detection kit based on Y-STR gene locus and Y-Indel as well as use method and application of fluorescence detection kit

A technology of Y-STR and y-indel, which is applied in the fields of biochemical equipment and methods, microbial measurement/inspection, etc., can solve the problems of high mutation rate of Y-STR locus, interference of family search, inaccurate test results, etc. Achieve the effects of shortening the amplification time, correct typing, and high specificity

Pending Publication Date: 2021-06-25
广州深晓基因科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

If the mutation rate of the Y-STR locus in the kit for constructing the Y database is high or the test results are inaccurate, individuals in the same family may also exhibit different haplotypes due to mutations, which may interfere with family search

Method used

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  • Fluorescence detection kit based on Y-STR gene locus and Y-Indel as well as use method and application of fluorescence detection kit
  • Fluorescence detection kit based on Y-STR gene locus and Y-Indel as well as use method and application of fluorescence detection kit
  • Fluorescence detection kit based on Y-STR gene locus and Y-Indel as well as use method and application of fluorescence detection kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0057] This embodiment provides a fluorescent detection kit, which includes the components shown in Table 3:

[0058] table 3

[0059]

[0060] Among them, the alleles and positive control typing information are shown in Table 4 below:

[0061] Table 4

[0062]

[0063]

[0064]

[0065] Wherein the primer pair can be designed according to the nucleotide sequence of each gene locus.

Embodiment 2

[0067] This example is used to identify whether the fluorescence detection kit provided in Example 1 is correct for typing each allele.

[0068] (1) Amplification: The reagents in the kit provided in Example 1 need to be completely dissolved before use, vortexed and oscillated to mix well, and different amplification systems are used according to the actual sample volume after brief centrifugation. The amplification system used in this example is shown in Table 5 below;

[0069] Wherein PCR reaction solution: primer pair mixture: amplification template=2:2:1;

[0070] table 5

[0071] ingredient name 10μL system PCR reaction solution 2μL primer pair mix 2μL Amplified template (0.1~2ng) 1μL Enzyme-free water Supplement to 10 μL

[0072] The amplification program was: pre-denaturation at 95°C for 5 min; denaturation at 95°C for 10 s, annealing at 62°C for 60 s, and extension at 70°C for 20 s, a total of 28 cycles; final extension at 6...

Embodiment 3

[0079] This embodiment is used to verify other relevant properties of the detection reagent provided in Example 1, and the detection method used is the same as that in Example 2.

[0080] (1) Fluorescence standard detection: Allelic genotyping and molecular weight standards of all loci were spectrally corrected with fluorescent standards, and when the peak height was below 2500 relative fluorescence intensity units, there was no peak height higher than 50 relative fluorescence intensity units. Penetration peak.

[0081] (2) Detection of molecular weight standards: The bands contained in the internal standard should be clear and distinct; the standard deviation from the allelic typing standard for repeated inspections is less than ±0.2bp: the peak height of each DNA fragment in the internal standard is greater than 500 Relative fluorescence intensity unit: The internal standard is detected by electrophoresis alone, and no non-specific bands appear.

[0082] (3) Positive DNA co...

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Abstract

The invention provides a fluorescence detection kit based on a Y-STR gene locus and Y-Indel as well as a use method and application of the fluorescence detection kit. The fluorescence detection kit can be used for simultaneously analyzing 38 Y-STR gene loci and 3 Y-indel in a single PCR. According to the kit, a multicolor fluorescence labeling composite amplification technology is adopted, amplification can be rapidly completed, a unique locus combination mode and a primer sequence better conform to the genetic characteristics of Chinese people, enough universal loci are reserved, exchange and sharing of DNA data are facilitated, and the kit better conform to the genetic characteristics of Chinese people; meanwhile, an amplification product is stable and long-acting, the Y chromosome specificity is high, complete male DNA typing can still be obtained under the high-background female DNA condition, and the method is suitable for extracting samples such as DNA, blood spots, saliva spots and oral swabs and can be used for multiple purposes such as paternal genetic relationship identification, individual recognition and public security library building.

Description

technical field [0001] The present invention relates to a genetic locus fluorescent marker compound amplification inspection system, in particular to a human DNA typing kit, in particular to a fluorescent detection kit based on Y-STR locus and Y-Indel and its use method and application. Background technique [0002] The Y chromosome is a sex chromosome, which can be divided into two regions according to different inheritance methods, namely the pseudoautosomal region (PAR) located at both ends of the Y chromosome and the male-specific region (MSY). The male-specific region is also known as the non-recombining region (NRY), which accounts for about 95% of the Y chromosome. It will not recombine with any chromosome and is strictly paternally inherited. The offspring of the same family have exactly the same Y chromosome Non-recombination regions (excluding mutations). [0003] Y-chromosome genetic markers, including short tandem repeats (STRs), single nucleotide polymorphisms ...

Claims

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Application Information

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IPC IPC(8): C12Q1/6888C12Q1/6858
CPCC12Q1/6888C12Q1/6858C12Q2600/156C12Q2563/107C12Q2531/113
Inventor 文少卿徐旭鼎林大钧郑诚浩李元元雷波
Owner 广州深晓基因科技有限公司
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