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SN/T 2474-2010: Detection of Phytophthora sojae with real-time PCR
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SN/T 2474-2010: Detection of Phytophthora sojae with real-time PCR

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Detection of Phytophthora sojae with real-time PCR Exit inspection and quarantine industry standard book People's Republic of China Phytophthora real-time PCR pathogen detection Issued on. 2010-01-10 2010-07-16 implementation People's Republic of China The State Administration of Quality Supervision, Inspection and Quarantine released

Foreword

Please note that some content of this document may involve patents, publication of this document Institutions do not assume the responsibility to identify these patents. This standard is proposed and managed by the National Certification and Accreditation Administration Committee. This standard was drafted. People's Republic of China SZCIQ, Nanjing Agricultural University, the People's Republic of China Liaoning Entry Inspection and Quarantine Bureau, Chinese Academy of Inspection and Quarantine, People's Republic of China Shanghai Exit Inspection and Quarantine, People's Republic of China Exit Inspection and Quarantine of Heilongjiang, China Agricultural University. The main drafters. Zhang Guiming, Wang Yuan-chao, WU Ji Yun, Peng gold fire, Yan Jin, Cheng Yinghui, Wang Ying, Yi Jianping Cui Shu, Yang Weidong, Yang Liqun, Nie Wenge, Miao Jian Kun. This standard is the first release of the entry-exit inspection and quarantine industry standards. Phytophthora real-time PCR pathogen detection

1 Scope

Identification methods. This standard applies to soybeans mixed with the soil mass Phytophthora pathogen detection.

2 Normative references

The following documents contain provisions which, through reference in this standard and become the standard terms. For dated references, subsequent Amendments (not including errata content) or revisions do not apply to this standard, however, encourage the parties to the agreement are based on research Whether the latest versions of these documents. For undated reference documents, the latest versions apply to this standard. Laboratory use specifications and test methods GB/T 6682 Analysis SN/T 1131 Phytophthora quarantine and identification of bacteria Principle 3 3.1 taxonomic status Taxonomic position. algae bacteria community Chromista, Oomycota Oomycota, Pythium head Pythia1es, Pythium Branch Pythiaceae, Phytophthora 3.2 real-time PCR Fluorescence signal with an increase in PCR product and enhance the principle of the PCR amplification process, continuously detect the reaction body Changes in the fluorescence signal lines, according to the mean and standard fluorescent signal baseline average difference is greater than the average value obtained fluorescence threshold value that is close Enhanced fluorescence signal set to a predetermined number of PCR cycles when the threshold value, i.e., the Ct value. Ct value determination based on whether the sample with Phytophthora Germs.

4 sampling method

Soybeans mixed soil samples according to SN/T 1131 carried out.

5 Apparatus and reagents

5.1 The main instruments 5.1.1 oven. 5.1.2 Autoclave. 5.1.3 shaker. 5.1.4 water bath. 5.1.5 water meter. 5.1.6 High-speed refrigerated centrifuge. 5.1.7 real-time PCR instrument. 5.1.8 Micropipettes (0.1μL ~ 2.5μL, 2μL ~ 20μL, 20μL ~ 200μL, 100μL ~ 1000μL). 5.1.9 Vacuum drained instrument. ...