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GB/T 22916-2008 English PDF

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GB/T 22916-2008: Protocol of fluorogenic RT-PCR for vesicular stomatitis virus
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Basic data

Standard ID GB/T 22916-2008 (GB/T22916-2008)
Description (Translated English) Protocol of fluorogenic RT-PCR for vesicular stomatitis virus
Sector / Industry National Standard (Recommended)
Classification of Chinese Standard B41
Classification of International Standard 11.220
Word Count Estimation 9,971
Date of Issue 2008-12-31
Date of Implementation 2009-05-01
Regulation (derived from) National Standard Approval Announcement 2008 No.25 (Total No.138)
Issuing agency(ies) General Administration of Quality Supervision, Inspection and Quarantine of the People's Republic of China, Standardization Administration of the People's Republic of China
Summary This standard specifies the method of operation of vesicular stomatitis virus fluorescent RT-PCR detection. This standard applies to animals and their products in the detection of vesicular stomatitis virus.

GB/T 22916-2008: Protocol of fluorogenic RT-PCR for vesicular stomatitis virus

---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.
Protocol of fluorogenic RT-PCR for vesicular stomatitis virus ICS 11.220 B41 National Standards of People's Republic of China Vesicular stomatitis virus fluorescent RT-PCR detection method Posted 2008-12-31 2009-05-01 implementation Administration of Quality Supervision, Inspection and Quarantine of People's Republic of China Standardization Administration of China released

Foreword

The standard reference to the OIE (OIE) "Terrestrial Animals Manual of diagnostic tests and vaccines (mammals, birds and bees)" (5th edition). Appendix A of this standard is a normative appendix, Appendix B is an informative annex. The standard proposed by the People's Republic of China Ministry of Agriculture. This standard by the National Standardization Technical Committee on Animal Epidemic Prevention. This standard was drafted. People's Republic of China SZCIQ, People's Republic of China Yunnan Entry-Exit Inspection and Quarantine Bureau. The main drafters of this standard. flower Qun Yi, Zhou Xiaoli, Zeng Ling, Caochen Fu, Zhanai Jun, Zhang Caihong, Lin Qingyan, Chen Bing, Yang Yunqing, Sun Jie. Vesicular stomatitis virus fluorescent RT-PCR detection method

1 Scope

This standard specifies the method of operation of vesicular stomatitis virus fluorescent RT-PCR detection. This standard applies to animals and their products in detecting vesicular stomatitis virus.

2 Acronyms

The following abbreviations apply to this standard. 2.1 fluorescent RT-PCR Fluorescence reverse transcription - polymerase chain reaction. The number of cycles when fluorescence signal of each reaction tube reaches a set threshold experienced. 2.3 RNA RNA. 2.4 DEPC Dicarbonate phospholipids. 2.5 PBS Phosphate buffered saline (recipe see Appendix A). 2.6 T Rao enzyme q T Rao q DNA polymerase. 2.7 Vp Vp S Vesicular stomatitis virus. Principle 3 Vesicular stomatitis virus is a RNA virus, according to a specific gene sequence of a total of two types of vesicular stomatitis virus, A pair of specific primers And a fluorescent substance-specific dual-labeled probes. Through rigorous design and selection of primers and probes, covering two vesicular stomatitis virus Type and mutant strains of vesicular stomatitis virus-specific universal primers and probes. Probe 5 'end FAM labeled fluorescein fluorescence emitted by it Light can be received detection equipment, the report called fluorophores; 3 'TAMRA labeled fluorescein, which can absorb at close range within the 5' end report Fluorophore fluorescence signal emitted, called quenching fluorophore. When amplification, T Rao q enzyme plays its 5 '→ 3' exonuclease end functions Can the probe into a single nucleotide hydrolysis, eliminate obstacles, the report marks both ends of the fluorophore and quencher probe fluorophores are free in solution , The instrument detects the fluorescent signal emitted.

4 Materials and Reagents

4.1 Instruments and Equipment 4.1.1 RT-PCR fluorescence detector. 4.1.2 high-speed desktop refrigerated centrifuge (centrifugal speed 12000r/min or more). 4.1.3 tabletop centrifuge (centrifugal speed 3000r/min). 4.1.4 Mixer. 4.1.5 refrigerator (2 ℃ ~ 8 ℃ and -20 ℃ two kinds). 4.1.6 Micro adjustable pipette (5μL, 10μL, 100μL, 1000μL) and supporting filter tips.

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