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GB/T 40183-2021: Determination of DNA methylation - Pyrosequencing
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Determination of DNA methylation - Pyrosequencing
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GB/T 40183-2021
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Basic data | Standard ID | GB/T 40183-2021 (GB/T40183-2021) | | Description (Translated English) | Determination of DNA methylation - Pyrosequencing | | Sector / Industry | National Standard (Recommended) | | Classification of Chinese Standard | A21 | | Word Count Estimation | 6,665 | | Issuing agency(ies) | State Administration for Market Regulation, China National Standardization Administration | GB/T 40183-2021: Determination of DNA methylation - Pyrosequencing---This is a DRAFT version for illustration, not a final translation. Full copy of true-PDF in English version (including equations, symbols, images, flow-chart, tables, and figures etc.) will be manually/carefully translated upon your order.
Determination of DNA methylation-Pyrosequencing
ICS 07.080
A21
National Standards of People's Republic of China
DNA methylation determination by pyrosequencing
Released on 2021-05-21
2021-12-01 implementation
State Administration of Market Supervision and Administration
Issued by the National Standardization Management Committee
Foreword
This standard was drafted in accordance with the rules given in GB/T 1.1-2009.
This standard was proposed and managed by the China National Institute of Standardization.
Drafting organizations of this standard. Hebei Medical University, China National Institute of Standardization, Hebei Food Inspection and Research Institute, Chinese Academy of Sciences Genetics and Development
Agricultural Resources Research Center, Institute of Educational Biology, Hebei Normal University.
The main drafters of this standard. Lu Pin, Ma Aijin, Zhang Yan, Lin Hao, Li Yongpeng, Li Dongming, Huang Yun, Yang Hongchao.
DNA methylation determination by pyrosequencing
1 Scope
This standard specifies the method for the determination of DNA methylation by pyrosequencing.
This standard applies to the determination of DNA methylation in animal and plant tissues or cells cultured in vitro.
2 Normative references
The following documents are indispensable for the application of this document. For dated reference documents, only the dated version applies to this article
Pieces. For undated reference documents, the latest version (including all amendments) is applicable to this document.
GB/T 6682 Analytical laboratory water specifications and test methods
3 Terms and definitions, abbreviations
3.1 Terms and definitions
The following terms and definitions apply to this document.
3.1.1
DNA methylation
Under the catalysis of DNA methyltransferase, S-adenosylmethionine is used as the methyl donor to transfer the methyl group to the specific structure of the base.
process.
3.2 Abbreviations
The following abbreviations apply to this document.
DNA. deoxyribonucleic acid (deoxyribonucleicacid)
PCR. polymerase chain reaction (polymerase chain reaction)
4 Principle
After the DNA is vulcanized by bisulfate, the unmethylated cytosine in the DNA double strand is converted into uracil, and the uracil is converted by PCR.
Converted to thymine, quantitatively based on the ratio of cytosine and thymine at the corresponding methylation sites in the PCR product.
5 Reagents or materials
Unless otherwise specified, only use analytical reagents.
5.1 Water. GB/T 6682, Grade II water.
5.2 Animal genomic DNA extraction kit.
5.3 Plant genomic DNA extraction kit.
5.4 20pmol/μL pyrosequencing primer.
5.5 PCR kit.
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