GB/T 41689-2022 English PDF

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GB/T 41689-2022: Soil quality - Direct extraction of soil DNA
Status: Valid

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Basic data

Standard ID: GB/T 41689-2022 (GB/T41689-2022)
Description (Translated English): Soil quality - Direct extraction of soil DNA
Sector / Industry: National Standard (Recommended)
Classification of Chinese Standard: B10
Classification of International Standard: 13.080.01
Word Count Estimation: 14,168
Date of Issue: 2022-10-14
Date of Implementation: 2023-05-01
Issuing agency(ies): State Administration for Market Regulation, China National Standardization Administration

GB/T 41689-2022: Soil quality - Direct extraction of soil DNA

---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.
Soil quality -- Direct extraction of soil DNA ICS 13.080.01 CCSB10 National Standards of People's Republic of China A method for direct DNA extraction from soil quality soil samples (ISO 11063.2020, IDT) Published on 2022-10-12 2023-05-01 Implementation State Administration for Market Regulation Released by the National Standardization Administration directory Preface III 1 Scope 1 2 Normative references 1 3 Terms and Definitions 1 4 Principle 1 5 Test material 2 5.1 Soil samples 2 5.2 Chemicals 2 5.3 Buffers and reagents 3 6 Instrument 3 7 DNA extraction step 4 7.1 Preparation of soil samples 4 7.2 Mechanical and chemical cracking 4 7.3 Protein precipitation 4 7.4 Nucleic acid precipitation and washing 4 7.5 Nucleic acid storage 4 8 Quality and content assessment of soil DNA4 8.1 Quality and purity of soil DNA 4 8.2 Content of soil DNA 5 9 Verification of Extractor 5 10 Test report 5 Appendix A (Informative) Differences between this document and the method for direct DNA extraction from soil samples in ISO 11063.20126 Appendix B (Informative) Available Methods for Purification of Soil DNA Extracts7 Reference 8

foreword

This document is in accordance with the provisions of GB/T 1.1-2020 "Guidelines for Standardization Work Part 1.Structure and Drafting Rules of Standardization Documents" drafted. This document is equivalent to ISO 11063.2020 "Method for Direct DNA Extraction from Soil Quality Soil Samples". The following minimal editorial changes have been made to this document. ---The mass calculation of potassium acetate in 5.3.7 of ISO 11063.2020 is incorrect, and the value is corrected from 176.5 to 294.4. Please note that the content of this document may involve patents. The issuing agency of this document assumes no responsibility for identifying patents. This document is proposed by the Ministry of Agriculture and Rural Affairs of the People's Republic of China. This document is under the jurisdiction of the National Soil Quality Standardization Technical Committee (SAC/TC404). This document was drafted by. Nanjing Soil Research Institute, Chinese Academy of Sciences, Institute of Subtropical Agroecology, Chinese Academy of Sciences, Jiangsu Quality and Standardization Institute, etc. The main drafters of this document. Pan Xianzhang, Guo Zhiying, Lin Xiangui, Wei Wenxue, Xiao Tingting. A method for direct DNA extraction from soil quality soil samples

1 Scope

This document describes a method for the direct extraction of DNA from soil samples, and these extracted DNAs are used to Analysis of soil bacterial community abundance and composition by biological techniques [including quantitative real-time PCR (qPCR)]. This method is mainly suitable for agricultural and forest soil. This method may not be suitable for soils rich in organic matter (such as peat), soils heavily contaminated with organic pollutants or heavy metals. Direct DNA extraction from soil samples provides a unique perspective for studying microbial community alpha and beta diversity. by soil DNA Next-generation sequencing of PCR-amplified amplicons will facilitate the development of routine tools for microbial community monitoring in soil environments.

2 Normative references

The contents of the following documents constitute essential provisions of this document through normative references in the text. Among them, dated citations documents, only the version corresponding to that date applies to this document; for undated references, the latest edition (including all amendments) applies to this document. ISO 18400-206 Soil quality sampling Part 206.Soils for laboratory determination of microbial processes, biomass and diversity Guidelines for aerobic collection, handling and storage (Soilquality-Sampling-Part 206.Collection, handling and storage of thelaboratory) Note. GB/T 32725-2016 Guidelines for aerobic collection, treatment and storage of soils for laboratory determination of microbial processes, biomass and diversity (ISO 10381-6. 20091), IDT) 1) ISO 10381-6.2009 has been replaced by ISO 18400-206.2018.

3 Terms and Definitions

The following terms and definitions apply to this document. ISO and IEC maintain terminology databases for standardization at. 3.1 soil DNA soilDNA DNA extracted from living soil microorganisms and residual DNA of dead microorganisms.

4 Principles

Follow the extraction procedure below to directly extract DNA from 1 g soil samples (equivalent dry weight). Soil with extraction buffer and glass beads added Soil samples were subjected to mechanical and chemical lysis. Lysis steps, such as bead shaking, are also very useful for DNA extraction from difficult-to-lyse microorganisms. crucial step. Then, the samples were incubated at 70 °C for 30 min for chemical lysis. Centrifuge briefly to remove soil debris and collect supernatant liquid. Potassium acetate was added to the supernatant fraction to precipitate proteins. After centrifugation, the supernatant was collected again, and cold isopropanol was added to precipitate nucleic acids. centrifugal, so The nucleic acid precipitate was washed with 70% ethanol and dissolved in molecular biology grade ultrapure water or TE buffer. Electroporation through agarose gel ......

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