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GB/T 22333-2008 English PDF

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GB/T 22333-2008: Reverse transcription polymerase chain reaction for Japanese B encephalitis virus
Status: Obsolete
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PDF similar to GB/T 22333-2008


Standard similar to GB/T 22333-2008

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Basic data

Standard ID GB/T 22333-2008 (GB/T22333-2008)
Description (Translated English) Reverse transcription polymerase chain reaction for Japanese B encephalitis virus
Sector / Industry National Standard (Recommended)
Classification of Chinese Standard B41
Classification of International Standard 11.220
Word Count Estimation 6,640
Date of Issue 2008-08-22
Date of Implementation 2008-12-01
Regulation (derived from) National Standard Approval Announcement 2008 No.14 (Total No.127)
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 technical requirements for reverse transcription -polymerase chain reaction test of Japanese encephalitis virus nucleic acid. This standard is applicable to the detection of animal brain tissue of Japanese B encephalitis virus nucleic acid.

GB/T 22333-2008: Reverse transcription polymerase chain reaction for Japanese B encephalitis 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.
Reverse transcription polymerase chain reaction for Japanese B encephalitis virus ICS 11.220 B41 National Standards of People's Republic of China Japanese encephalitis virus Reverse transcription polymerase chain reaction test method Posted 2008-08-22 2008-12-01 implementation Administration of Quality Supervision, Inspection and Quarantine of People's Republic of China Standardization Administration of China released

Foreword

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. the Ministry of Agriculture veterinary diagnostic center. The main drafters of this standard. Wang Hongwei, Sun Ming, Wang Chuanbin, Zhao Chen Xi, TIAN Ke-gong. Japanese encephalitis virus Reverse transcription polymerase chain reaction test method

1 Scope

This standard specifies the technical requirements of reverse transcription polymerase chain reaction test of Japanese encephalitis virus nucleic acid. This standard is applicable to the detection of animal brain tissue Japanese encephalitis virus nucleic acid.

2 Reagents

2.1 denaturing solution (see Section A.1 chapter). 2.2 2mol/L sodium acetate (pH4.0). 2.3 phenol - chloroform - isoamyl alcohol mixture (see Section A.2 chapter). 2.4 2.5mmol/LdNTP. 2.5 8mmol/L on the downstream primer mixture (primer sequences see section A.3 chapter). 2.6 10x PCR buffer. 2.7 1% ethidium bromide (EB) solution. 2.8 TAE electrophoresis buffer. 2.9 1% agarose gel. 2.10 loading buffer (see Chapter A.4). 2.11 Other reagents. 10U/μLAMV reverse transcriptase, 100bpDNA molecular mass standard, 50U/μLRNA inhibitors, 5U/μLT Rao q DNA polymerase, isopropanol, 75% ethanol solution, 25mmol L magnesium chloride solution /, DEPC-treated sterile double Distilled water.

3 laboratory conditions

3.1 laboratory level requirements Laboratories must biosafety level Ⅲ (BSL-3 level) laboratories. 3.2 Laboratory partition RT-PCR test-area reaction liquid preparation area (dosing area), nucleic acid extraction zone, the amplified region, zone electrophoresis. Flow sequence for the dosing area → Amplified nucleic acid extraction District → District → electrophoresis area. Prohibited equipment and reagents back. 3.3 laboratory should be equipped with equipment and supplies 3.3.1 Instrument. analytical balance, high-speed centrifuge, a vacuum dryer, PCR amplification device, electrophoresis, electrophoresis tank, UV gel imager (or purple Outside the analyzer), liquid nitrogen tanks or -70 ℃ refrigerator, microwave, tissue grinder, -20 ℃ refrigerator, water bath, adjustable pipette (maximum range points Do as 2μL, 20μL, 200μL, 1000μL). 3.3.2 supplies. scissors ophthalmology, ophthalmic tweezers, weighing paper, 10mL disposable syringe, 1.5mL sterile centrifuge tube, 0.2mL thin wall PCR Pipe, agarose, 500mL graduated cylinder, 500mL conical flask, pipette tips (10μL, 200μL, 1000μL), sterilized double-distilled water. Procedure 4 Sample collection and processing 4.1 4.1.1 Sample collection. the dying animals, culling of animals or aborted fetus brain tissues. 4.1.2 Sample treatment. The samples were each treated.

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