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

Method for detecting gene promoter methylation

A detection method, a methylation technology, applied in the fields of life science-biochemistry and molecular biology, which can solve the problems of difficult repeatability of methylation detection, low amplification efficiency and MSP

Inactive Publication Date: 2008-02-27
冯景 +1
View PDF0 Cites 7 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] The present invention proposes a method for detecting methylation of gene promoters to solve the problems of repeated methylation detection, sodium bisulfite modification, multi-promoter methylation, multiple methylation-specific PCR amplification, and low amplification efficiency. Questions about reacting with MSP

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

Examples

Experimental program
Comparison scheme
Effect test

Embodiment Construction

[0010] (1). Sodium bisulfite modification

[0011] 1.1 Reagent preparation and modification process

[0012] 1.1.1 Reagent preparation Hydroquinone 55mg in 5ml ddH 2 In O, 5.2g of sodium bisulfite was dissolved in 10ml of water and about 1.4ml of 3M sodium hydroxide was used to accurately adjust the pH to 5.0 (prepared before use). The modified concentration is above 3.0mol / L to ensure thorough modification.

[0013] 1.1.2 Modification process Take 1 μg of DNA, add double distilled water to 50 μl, add 3M NaOH, 5.5 μl, and put it at 54°C for 20 minutes to denature. Add 30 μl of 10 mmol / L hydroquinone (prepared just before use) and 520 μl of 3.9M sodium bisulfite (pH 5.0) (prepared just before use) to the denatured DNA, and incubate at 54° C. for 16 hours. Then purify with a purification column, add 50 μl of pre-warmed 80°C double distilled water to dissolve the purified DNA, add 5.5 μl of 3M NaOH, let stand at 25 degrees for 15 minutes, add 56 μl of 10M NH4Ac to stand at roo...

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 present invention relates to a detection method of gene promoter methylation. Said method includes the following steps: firstly, using freshly-prepared modification reagent to modify extracted DNA, after the modified DNA is passed through column, making purification and desulfurization reaction, then using absolute ethyl alcohol and salmon sperm DNA to enrich micro modified DNA, further selecting and using specific PCR GC Buffer to make multiple methylation specific PCR amplification, then adopting electrophoresis technique to analyze result.

Description

technical field [0001] The invention belongs to the field of life science-biochemistry and molecular biology, and relates to a method for detecting gene promoter methylation. Background technique [0002] DNA methylation is an enzyme-mediated chemical modification process in higher vertebrate organisms, which affects gene transcription and expression, and is closely related to tumorigenesis. Changes in DNA methylation are a key factor in tumorigenesis and one of the inactivation methods of tumor suppressor genes. It leads to abnormal expression of genes related to cell proliferation and differentiation through genetic and epigenetic mechanisms, resulting in cell loss. Normal gene regulation results in malignant transformation to form tumors. At present, there are not many methods for detecting DNA methylation status. Methylation-specific PCR (methylation-specific PCR, MSP) is a fast and sensitive method for gene methylation analysis[2]. It can detect tumor cells in which 1...

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
IPC IPC(8): C12Q1/68
Inventor 冯景吕军王玉平张吉才
Owner 冯景
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