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High-resolution melting curve technology-based tardive deafness gene mutation detection primer group, kit and application

A delayed-onset deafness, high-resolution melting technology, applied in the field of molecular biology technology and medical testing, can solve the problem that it is not suitable for the detection of a single or a small number of SNP loci, cannot detect unknown SNP loci, and cannot process a large number of samples at the same time, etc. problem, to achieve the effect of high specificity, easy operation and low cost

Inactive Publication Date: 2017-09-08
THE FIRST AFFILIATED HOSPITAL OF ZHENGZHOU UNIV
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Problems solved by technology

The disadvantage is that it is only suitable for detecting known SNPs, but cannot detect unknown SNP sites; and the synthesis cost of Taqman probes is high, which is not an economical choice for the detection of small or medium-throughput samples; in addition, this method Due to the limitation of the sequence near the SNP site, some SNPs are not suitable for Taqman probes
Gene chip technology, compared with traditional instrument detection methods, has the characteristics of automation, high throughput, miniaturization, and easy operation. The disadvantage is that it needs special scanning equipment, which is expensive and not suitable for the detection of single or small SNP sites. , and cannot handle a large number of samples at the same time

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  • High-resolution melting curve technology-based tardive deafness gene mutation detection primer group, kit and application
  • High-resolution melting curve technology-based tardive deafness gene mutation detection primer group, kit and application
  • High-resolution melting curve technology-based tardive deafness gene mutation detection primer group, kit and application

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Embodiment Construction

[0046] Preferred embodiments of the present invention will be described in more detail below with reference to the accompanying drawings.

[0047] 1. Primer synthesis

[0048] Entrust Shanghai Biotechnology Co., Ltd. to synthesize primers with the base sequences shown in SEQ ID NO: 1-SEQ ID NO: 10.

[0049] 2. Extract the genomic DNA from the peripheral blood or plantar blood samples and perform PCR amplification.

[0050] For the PCR experiment, Tiangen Biochemical Technology High Affinity HotStart Taq Kit (ET108) was selected. In the PCR reaction system, add buffer, dNTPs, DNA polymerase, primers, template DNA, H 2 O. In order to reduce human error and simplify the operation process, in this embodiment, the PCR amplification reagents are first configured as Mix (including HA Buffer, dNTPs, H 2 O, LC GreenPlus, DNA Polymerase), and then mixed with specific primer pairs of different mutations respectively, and aliquoted into each reaction well; finally, different sample DN...

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Abstract

The invention relates to the fields of a molecular biological technique and medical examination, in particular to a high-resolution melting curve technology-based tardive deafness gene mutation detection primer group, a kit and application. The primer group comprises primers with base sequences as shown in SEQ ID NO:1 to SEQ ID NO:8. The kit provided by the invention is used for performing detection, and is simple and convenient to operate, high in specificity, high in result repeatability and low in cost.

Description

technical field [0001] The present invention relates to the field of molecular biology technology and medical examination, more specifically, relates to the late-onset deafness gene mutation detection primer set based on high-resolution melting curve technology, and a kit including the primer set, as well as the above primer set and kit Application as a detection reagent for delayed deafness gene mutation. Background technique [0002] Deafness is the most common disease that causes communication disorders, and it is also one of the most common clinical genetic diseases, which seriously affects the quality of life of human beings. The current situation of deaf people in my country is particularly serious. According to the statistics of the second (2006) national sample survey of disabled persons, the number of people with hearing and language disabilities in my country is as high as 27.8 million, accounting for 34% of the total number of disabled people, of which the number ...

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6858C12Q1/6883C12Q2600/156C12Q2527/107C12Q2531/113C12Q2563/107
Inventor 梅世月孔祥东白楠亢鸿飞
Owner THE FIRST AFFILIATED HOSPITAL OF ZHENGZHOU UNIV
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