Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Reagent case for detecting POU3F4 gene 499C>T mutation

A technology of POU3F4 and detection reagents, which is applied in the direction of microbial measurement/inspection, biochemical equipment and methods, etc., and can solve the problems of limited research methods, deep inner ear, slow progress, etc.

Inactive Publication Date: 2008-08-27
GENERAL HOSPITAL OF PLA
View PDF0 Cites 10 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Hereditary deafness has been documented in the 16th century, but due to the deep inner ear, small size, and limited research methods, the progress in understanding and understanding the genes that control the auditory system has been slow
There is no report on the discovery of the 499C>T mutation of the POU3F4 gene in deaf patients

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Reagent case for detecting POU3F4 gene 499C>T mutation
  • Reagent case for detecting POU3F4 gene 499C>T mutation
  • Reagent case for detecting POU3F4 gene 499C>T mutation

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0052] 1. Preparation of blood sample DNA of the subject to be tested

[0053] 1. Research object

[0054] A Chinese family with X-linked genetic congenital severe hearing loss, the patient showed congenital severe hearing loss, temporal bone CT examination showed enlarged internal auditory canal, showing an X-linked recessive inheritance pattern. A total of 17 family members were investigated, including 10 males and 7 females. There were 2 patients, both male, with the same audiological phenotype. According to the following method, 8 of them, including 5 females and 3 males, were tested for the POU3F4 gene, and it was found that 2 male patients were hemizygous for the mutation, and the other normal male gene was wild type, and a total of 5 female patients were detected The 3 heterozygous carriers were the mother, grandmother and aunt of the proband. No mutations were found in the other two women. In addition, 110 normal controls with normal hearing and no genetic backgrou...

Embodiment 2

[0087] 1. Purification of PCR products——96-well plate method

[0088] 1. Add 50 μl sterile water to the 96-well plate containing the PCR product and mix well.

[0089] 2. Transfer it to the Millipore purification plate, put it on the vacuum pump for about 3 minutes, and see that there is no water in the purification plate.

[0090] 3. Add 50 μl of deionized water to the purification plate again, and continue to filter until there is no water in the purification plate.

[0091] 4. Remove the purification plate from the vacuum pump, add 20 μl of deionized water to the plate, let it rest for 15 minutes, shake it for another 15 minutes, and then suck it into a new 96-well plate.

[0092] 5. Store in a -20°C refrigerator.

[0093] 2. Quantification by electrophoresis

[0094] 1. Sample preparation

[0095] Take a 96-well spotting plate, add 6 μl of sample buffer to each well, remove the PCR product (2 μl) from each well of the cavity plate containing the PCR product, transfer to ...

Embodiment 3

[0108] 1. Purity and dosage requirements of PCR product DNA template

[0109] DNA purity: OD 260 / OD 280 = 1.6 to 2.0.

[0110] DNA concentration: PCR product 10ng / μl.

[0111] DNA consumption:

[0112] PCR product

[0113] 100-200bp 1-3ng

[0114] 200-500bp 3-10ng

[0115] 500-1000bp 5-20ng

[0116] 1000-2000bp 10-40ng

[0117] >2000bp 40-100ng

[0118] 2. Sequencing reaction

[0119] 1. The reagents required for the sequencing reaction should be freshly prepared, and the reagents that need to be sterilized by autoclaving must be sterilized before use. The equipment required for the sequencing reaction (such as 384-well plates, tips, etc.) should also be clean and sterile.

[0120] 2. In order to ensure the freshness of sequencing samples and reaction reagents, it should be operated on ice when adding samples.

[0121] 3. The current reaction system is 5 μl, and the amount of various reagents added is shown in Table 2.

[0122] Table 2...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

The invention relates to POU3F4 mutant gene which has 499C> T mutation, which is relative to human genetic deafness. The invention provides a reagent kit for detecting POU3F4 gene 499 C>T mutation, and a detection process which is used to diagnose the reason and kind of deafness generation through detecting whether patients have the mutation genes. The mutant gene and the detection process are beneficial for the POU3F4 mutation screening work of deafness patients which is carried out on clinics, and provides services for the diagnosis and treatment of deaf patients.

Description

technical field [0001] The invention relates to a kit for detecting the 499C>T mutation gene of POU3F4 gene. [0002] The invention also relates to a new POU3F4 mutant gene and its application in diagnosing and / or treating human hereditary deafness. Background technique [0003] Hereditary deafness is divided into nonsyndromic hearing impairment (NSHI) and syndromic hearing impairment (SHI). All NSHI and most SHI are Mendelian monogenic diseases, and very few SHI are chromosomal diseases. Deafness is the most common cause of communication impairment. It is estimated that about 700 million people in the world have a hearing loss of at least 55dB. Among them, prelingual deafness is a common form of onset, and it is also a category that has a serious social impact. The incidence rate is 1 / 1000, and about half of them are caused by genetic factors. This population has exceeded 27 / 10,000, of which 70% are NSHI and 30% are SHI. Inherited deafness has been documented in the ...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
Inventor 王秋菊赵亚丽韩东一韩明鲲
Owner GENERAL HOSPITAL OF PLA
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com