Compound amplification detection kit containing forty InDel genetic polymorphic sites of human X chromosome

A genetic polymorphism and detection kit technology, applied in the field of kits for detecting human X chromosome InDel genetic polymorphism sites, can solve the problem that the number of systems and products is small, and the number of detection sites has no great advantage, etc. problems, to achieve the effect of good sample compatibility, good typing efficiency, and low mutation rate

Active Publication Date: 2019-12-17
GUANGDONG HUAMEI ZHONGYUAN BIOLOGICAL SCI & TECH +2
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AI Technical Summary

Problems solved by technology

[0004] Although there have been some reports on the application of X-InDel genetic markers at home and abroad, the number of systems and products that can be applied to actual identification cases is still small; There is no great advantage in the number of sites, so it is still necessary to develop a polymorphic genotyping system that includes more detection sites, stronger discrimination ability, and better stability

Method used

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  • Compound amplification detection kit containing forty InDel genetic polymorphic sites of human X chromosome
  • Compound amplification detection kit containing forty InDel genetic polymorphic sites of human X chromosome
  • Compound amplification detection kit containing forty InDel genetic polymorphic sites of human X chromosome

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] Example 1: A multiple amplification detection kit comprising 40 InDel genetic polymorphism sites of human X chromosome.

[0037] 1) Design of primers

[0038] 1-1) Screening of 40 X-InDel polymorphic sites

[0039] Based on the dbSNP database established by the National Center for Biotechnology Information (NCBI), combined with relevant literature reports, and according to the proposed screening conditions for InDel polymorphic sites, 40 polymorphic sites were screened from the X chromosome with a length of about 150 Mb. InDel polymorphic loci ensure that they are nearly evenly distributed on the X chromosome with a considerable distance between them. On this basis, Amelogenin was added as a sex identification site. The specific screening criteria for X-InDel loci are as follows: ①The length difference of X-InDel allelic fragments (the number of bases inserted or deleted) is between 3 and 30 bp; ②It is located in the intron region of X chromosome; ③dbSNP database The m...

Embodiment 2

[0055] Example 2: Sensitivity test of the multiple amplification detection kit containing 40 InDel genetic polymorphism sites of human X chromosome.

[0056] The sensitivity test was performed with the multiplex amplification detection kit described in Example 1. When preparing the amplification system, the template amounts of standard genomic DNA were set to 1ng, 0.5ng, 0.25ng, 0.125ng, 62.5pg, and 31.25pg, respectively. Amplification detection was performed according to the amplification system and procedure described in Example 1. The test results obtained as figure 2 Shown: When the amount of template is higher than or equal to 62.5pg, each detection site can produce peaks normally, and the balance of peak height between colors is good; when the amount of template is further reduced to 31.25pg, the two positions of rs16637 and rs36094418 The allele peak height of the point is lower than the analysis threshold; the sensitivity of the effective detection sample of the kit ...

Embodiment 3

[0057] Example 3: Identification of grandmother-mother-granddaughter relationship.

[0058] The sample source was provided by volunteers, and the sample type was saliva spot; DNA was extracted by chelex-100 method. Take 2×1.2mm saliva spots, add 80μL of 5% chelex-100 solution, incubate at 56°C for 30min, shake fully, keep at 95°C for 8min, shake and mix well, and centrifuge at 12000rpm to get the supernatant.

[0059] Amplification detection: Carry out according to the amplification conditions and detection conditions described in Example 1.

[0060] The amplified products were detected by capillary electrophoresis detection and analysis system, and the results were analyzed by data analysis software GeneMapper® ID-X.

[0061] Typing map such as image 3 As shown, a, b, and c in the figure are the X-InDel of the grandmother, mother, and granddaughter respectively; the specific typing results are as follows:

[0062]

[0063]

[0064] The results showed that the grandd...

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Abstract

The invention discloses a compound amplification detection kit containing forty InDel genetic polymorphic sites of human X chromosome. The forty InDel sites of the X chromosome and an individual identification location point are amplified simultaneously by a primer group. Compared with a traditional STR genetic marker, an insertion deletion genetic marker used in the compound amplification detection kit has the advantages of low mutation rate and high sensitivity; and the polymorphism detection sites involved in the compound amplification detection kit combine the advantages of an InDel genetic marker and the special genetic characteristics of the X chromosome, and effective supplementary means can be provided for identification of difficult or special cases.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a kit for detecting InDel genetic polymorphism site of human X chromosome. Background technique [0002] Short tandem repeats (STR) typing is a very important technical means in the field of forensic science, and plays an indispensable role in the construction of DNA databases, individual identification and paternity testing. However, with the application of STR typing technology more and more widely, some technical defects of its own have gradually become prominent. For example, the high mutation rate of STR loci has attracted more and more attention. This phenomenon is not conducive to the interpretation of paternity test results; secondly, the amplification product fragments of STR typing are generally long, which is not conducive to the detection and analysis of degraded samples; moreover, the number of STR loci with forensic application value is still limited, which is not condu...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6888
CPCC12Q1/6888C12Q2600/156
Inventor 刘超郑阳阳杜蔚安刘长晖郑文彦徐曲毅郑卫国陈林丽
Owner GUANGDONG HUAMEI ZHONGYUAN BIOLOGICAL SCI & TECH
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