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GB/T 22917-2008 English PDF

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GB/T 22917-2008: Protocol of fluorogenic RT-PCR for swine vesicular disease virus
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Basic data

Standard ID GB/T 22917-2008 (GB/T22917-2008)
Description (Translated English) Protocol of fluorogenic RT-PCR for swine vesicular disease virus
Sector / Industry National Standard (Recommended)
Classification of Chinese Standard B41
Classification of International Standard 11.220
Word Count Estimation 12,121
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 swine vesicular disease virus fluorescent RT-PCR detection. This standard applies to the animals and their products in the detection of swine vesicular disease virus.

GB/T 22917-2008: Protocol of fluorogenic RT-PCR for swine vesicular disease 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 swine vesicular disease virus ICS 11.220 B41 National Standards of People's Republic of China Swine vesicular disease 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. 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, Lu Kang body, Lv Jianjiang, Ruanzhou Xi, Yang Yunqing, Zhou Xiaoli, Yang Su, Dong Jun, Chenshu Kun. Swine vesicular disease virus fluorescent RT-PCR detection method

1 Scope

This standard specifies the method of operation swine vesicular disease virus fluorescent RT-PCR detection. This standard applies to animals and their products in the detection of swine vesicular disease 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 S D Vp Vp Swine vesicular disease virus. Principle 3 According to a specific gene sequence of swine vesicular disease virus synthesized a pair of specific primers and a dual-labeled probes specific fluorescence. Primers And probe design and through rigorous screening to cover all reported strains of swine vesicular disease virus. 5 'end fluorescent probes labeled FAM Fluorescein, 3 'TAMRA labeled fluorescein, which can absorb at close range within the 5' fluorophore reporter fluorescent signal emitted. Amplification, Since T Rao q enzyme 5 '→ 3' exonuclease activity, when extended to the fluorescent probe, which was cut to two groups separated, quenching effect disappears, fluorescent letter Number generation. Thus, by detecting the fluorescence signal is detected on a nucleic acid template.

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). Palm Desktop 4.1.3 or centrifuge 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. 4.1.7 1.5mL, 0.5mL siliconized Eppendorf tube. the Eppendorf tubes and emitters containing 0.1% DEPC soaked in distilled water three Overnight, 121 ℃ ± 2 ℃ autoclave 15min, 40 ℃ drying spare. Commercially available Eppendorf tubes and emitters have been suicide, can be directly

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