Deafness-related gene capture kit and application thereof

A technology of gene capture and kit, which is applied in the direction of DNA/RNA fragments, microbial determination/inspection, recombinant DNA technology, etc., and can solve the problems of few detection sites, expensive results, instability, etc.

Active Publication Date: 2020-07-07
迈基诺(重庆)基因科技有限责任公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Although each has its own advantages, it also has certain disadvantages, such as few detecti

Method used

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  • Deafness-related gene capture kit and application thereof
  • Deafness-related gene capture kit and application thereof
  • Deafness-related gene capture kit and application thereof

Examples

Experimental program
Comparison scheme
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Embodiment 1

[0051] Example 1. Preparation of deafness-related gene capture kit and special probes thereof

[0052] 1. Design and preparation of deafness-related gene capture probes

[0053] According to the whole exon sequence of deafness-related genes, a 120bp probe sequence was designed for the non-repeated region in each region, and each sequence was designed by moving along the gene position. Using in situ synthesis technology, a large number of designed probes were synthesized.

[0054] A large amount of biotin-labeled probes were amplified by PCR, and the specific method was as follows:

[0055] The probe synthesized above was dissolved in a total volume of 1.2ml of ddH 2 In O, take 5 μl of it as a DNA template, use general-purpose PCR primers (the sequence at the 5' end is GACTACATGGGACAT, and the sequence at the 3' end is GGAACCTACGACGTA), and divide into three tubes for PCR amplification, wherein the primer GACTACATGGGACAT is a primer labeled with biotin (Sequence 114, Sequenc...

Embodiment 2

[0129] Embodiment 2, the application of kit

[0130] The present invention will be described in further detail below through specific embodiments

[0131] 1. Kit capture detection of a deafness-related gene

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Abstract

The invention discloses a deafness-related gene capture kit and an application thereof. The invention provides a capture probe. The capture probe is as shown in 1) or 2) which are described in the specification, wherein 1) the capture probe is composed of probes as shown in sequences 1 to 113 in a sequence table; 2) the capture probe is composed of derivatives of probes in 1); and the derivative of each probe is a probe which is obtained by substituting and/or deleting and/or adding one or more nucleotides to each probe in 1) and has the same function. According to the deafness-related gene capture kit provided by the invention, target region sequencing analysis is carried out on a whole exon group or a deafness-related genome (respectively accounting for 1% and 0.01% of the whole genome)of a hereditary deafness patient; and most pathogenic mutation information of the disease is captured.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a deafness-related gene capture kit and its application. Background technique [0002] Deafness is one of the most common genetically heterogeneous diseases. Most hereditary deafness is a single-gene disease, and only a small number of deafness is caused by a combination of multiple genes. Among all deaf patients, hereditary deafness accounts for about 1 / 2, and there is about 1 case of deafness in every 500 newborns. According to whether the patient is accompanied by other organ diseases, it can be divided into syndromic deafness and non-syndromic deafness. Non-syndromic deafness can be divided into autosomal dominant inheritance, autosomal recessive inheritance, X-linked and Mitochondrial maternal inheritance. The main related genes include GJB2, GJB6, GJB3, SLC26A4, 12S rRNA, MYOTA, OTOF, among which the most common deafness-causing genes in China are GJB2, SLC26A4 an...

Claims

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Application Information

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IPC IPC(8): C12Q1/6883C12N15/11
CPCC12Q1/6883C12Q2600/156
Inventor 伍建姬晓雯王海丽王彩澜
Owner 迈基诺(重庆)基因科技有限责任公司
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