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A method for screening genes related to DNA methylation modification

A methylation and screening technology, applied in the field of DNA detection, can solve problems such as unknown gene functions, and achieve the effects of preventing DNA renaturation, improving sensitivity, and ensuring specificity

Inactive Publication Date: 2019-01-25
SHANXI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] With the advancement of technology, we can obtain more and more mutant materials, which facilitates the study of gene function, but the function of many genes is still unknown

Method used

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  • A method for screening genes related to DNA methylation modification
  • A method for screening genes related to DNA methylation modification
  • A method for screening genes related to DNA methylation modification

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Example 1: Screening of genes related to DNA methylguanine (M1G) modification in yeast

[0029] Take the cells of Saccharomyces cerevisiae wild-type and gene deletion mutant rad2 strains, respectively insert them into YPD liquid medium for activation overnight, then culture them at 30°C and 200-250r / min, take a small amount of culture medium every 2 hours, and detect OD 600 Value, draw the growth curve of wild-type and mutant bacteria, and count the cell concentration with a hemocytometer, find out the cell concentration and OD during the growth of the specific strain of cells 600 The value is linearly corresponding to the interval;

[0030] Cells in the logarithmic phase of growth were taken, and the drug treatment group was treated with peroxynitrite at a final concentration of 50 μmol / L, while the control group was treated without peroxynitrite; 30 ° C, 200 r / min for 3 hours;

[0031] Use the QIAGEN Genomic DNA Extraction Kit and operate according to the QIAGEN Gen...

Embodiment 2

[0044] Example 2: Screening of genes related to DNA methylguanine (M1G) modification in yeast

[0045] Take the cells of Saccharomyces cerevisiae wild type and gene deletion mutant apn1 strain, respectively insert them into YPD liquid medium to activate overnight, then culture them at 30°C, 200-250r / min, take a small amount of culture medium every 2 hours, and detect OD 600 Value, draw the growth curve of wild-type and mutant bacteria, and count the cell concentration with a hemocytometer, find out the cell concentration and OD during the growth of the specific strain of cells 600 The value is linearly corresponding to the interval;

[0046] Cells in the logarithmic phase of growth were taken, and the drug treatment group was treated with peroxynitrite at a final concentration of 50 μmol / L, while the control group was treated without peroxynitrite; 30 ° C, 200 r / min for 3 hours;

[0047] Use the QIAGEN Genomic DNA Extraction Kit and operate according to the QIAGEN Genomic DN...

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Abstract

The invention provides a method of screening DNA methylated modification related genes and belongs to the technical field of DNA detection. The method includes: utilizing a single-gene deletion type mutant biological material; adopting a chemiluminescence enzyme-linked immunoassay method to detect base methylation level in genome DNA strands; by comparing gene deletion type mutants with methylation level of genome DNA in normal control cells homologous with the gene deletion type mutants, analyzing relation between deleted genes in the mutants and DNA base methylated modification; screening genes related to DNA base methylated modification. The method has detection specificity on methylated bases in DNA and extremely high detection sensitivity. The method is simple and convenient in operation step, low in cost, high in stability, needless of large expensive equipment and capable of detecting multiple samples at the same time.

Description

Technical field: [0001] The invention relates to the technical field of DNA detection, in particular to a method for screening genes related to DNA methylation modification. technical background: [0002] Organisms will produce oxidative DNA damage during aerobic activity, and can activate the repair system in the body to repair DNA damage in time to maintain the stable genetic structure and normal function of the body. When the body's related gene function is missing or abnormal, the body's ability to repair damage is reduced. Due to the existence of gene complementation in organisms, many single-gene deletion mutants can maintain a physiological state under normal conditions, but when encountering environmental stimuli, the function of the mutants to maintain body homeostasis is low, which is different from the normal control group. [0003] The methylation modification of bases in DNA molecules is related to DNA oxidative damage, and also involves the latest epigenetic c...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6858
CPCC12Q1/6804C12Q2545/113
Inventor 仪慧兰仪民
Owner SHANXI UNIV