GB/T 40183-2021 in English
VALIDDetermination of DNA methylation—Pyrosequencing
- Issued on:2021-05-21
- Implemented on:2021-12-01
- File Format:PDF
- Delivery:Via email within 1~3 business days
$97.00
《GB/T 40183-2021DNA甲基化的测定 焦磷酸测序法》由424-cnis（中国标准化研究院）归口，主管部门为国家市场监督管理总局。
Introduction
Background and significance of the standard
As the first national standard in my country specifically for the determination of DNA methylation by pyrophosphate sequencer, this standard fills the gap in methodological standards in the field of epigenetic research. The standard took three years to formulate and integrated the research results of Hebei Medical University and other institutions in the fields of tumor epigenetics and agricultural breeding.
Analysis of core principles
Based on the bisulfite conversion-pyrophosphate sequencing technical route:
- Unmethylated cytosine (C) is converted into uracil (U) by bisulfite treatment
- U is converted into thymine (T) during PCR amplification
- Methylation quantification is achieved by detecting the C/T ratio through a pyrophosphate sequencer
Comparison of key experimental requirements
| Projects | Requirements of this standard | Differences in conventional methods |
|---|---|---|
| DNA starting amount | 10-20ng | Conventionally 50-100ng |
| Transformation efficiency | ≥98% | Conventional method 90-95% |
| Primer design | Need to avoid CpG islands | No special restrictions |
Implementation points
Typical application scenarios
In the study of early cancer screening, this standard was used to detect the methylation level of the RASSF1A gene promoter region:
- Tissue samples were treated with the Animal Genomic DNA Extraction Kit
- PCR amplification after bisulfite conversion
- Pyrosequencing showed the methylation rate of the gastric cancer group was (72.3±8.1)% vs. the normal group (4.2±2.7)%
Technology evolution analysis
Compared with the 2015 version of the industry standard, this version has the following major upgrades:
- Added Plant Genomic DNA extraction specifications
- Optimized the PCR system to 50μL (originally 100μL)
- Clearly specify the detailed parameters such as the amount of magnetic beads used (3μL)
Loading PDF document...
Error loading PDF. Please make sure the file is valid and try again.
We also recommend
-
GB/T 38475-2020 in English
Detection of mycotoxins in pigment—Rapid quantitative method of colloidal gold strip test
2020-11-19 -
GB/T 34800-2017 in English
Detection method of activity and impurity of proteinase K
2017-11-01 -
GB/T 40186-2021 in English
Determination of genetic material damage strength for microbial mutation breeding—Umu method
2021-05-21 -
GB/T 38790.1-2020 in English
Efficacy evaluation of bioactive peptides—Part 1:General principles
2020-04-28
