GB/T 36820-2018 English PDF

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GB/T 36820-2018: Detection method of Sugarcane streak mosaic virus by real-time reverse transcription polymerase chain reaction (RT-PCR)
Status: Valid

Standard ID	USD	BUY PDF	Lead-Days	Standard Title (Description)	Status
GB/T 36820-2018	199	Add to Cart	3 days	Detection method of Sugarcane streak mosaic virus by real-time reverse transcription polymerase chain reaction (RT-PCR)	Valid

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Basic data

Standard ID: GB/T 36820-2018 (GB/T36820-2018)
Description (Translated English): Detection method of Sugarcane streak mosaic virus by real-time reverse transcription polymerase chain reaction (RT-PCR)
Sector / Industry: National Standard (Recommended)
Classification of Chinese Standard: B16
Classification of International Standard: 65.020.01
Word Count Estimation: 10,176
Date of Issue: 2018-09-17
Date of Implementation: 2019-04-01
Issuing agency(ies): State Administration for Market Regulation, China National Standardization Administration

GB/T 36820-2018: Detection method of Sugarcane streak mosaic virus by real-time reverse transcription polymerase chain reaction (RT-PCR)

---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.
Detection method of Sugarcane streak mosaic virus by real-time reverse transcription polymerase chain reaction (RT-PCR) ICS 65.020.01 B16 National Standards of People's Republic of China Sugarcane striped mosaic virus real-time fluorescent reverse transcription polymerase Chain reaction (RT-PCR) detection method Published on.2018-09-17 Implementation of.2019-04-01 State market supervision and administration China National Standardization Administration issued

Foreword

This standard was drafted in accordance with the rules given in GB/T 1.1-2009. This standard is proposed and managed by the National Technical Committee for Phytosanitary Standardization (SAC/TC271). This standard was drafted. National Sugarcane Engineering Technology Research Center of Fujian Agriculture and Forestry University, and quality supervision and inspection of sugarcane and products of the Ministry of Agriculture. Center, Key Laboratory of Sugarcane Biology and Genetic Breeding, Fujian Province. The main drafters of this standard. Gao Sanji, Huang Meiting, Fu Huaying, Sun Shengren, Zhang Huili, Wang Jinda, Chen Rukai. Sugarcane striped mosaic virus real-time fluorescent reverse transcription polymerase Chain reaction (RT-PCR) detection method

1 Scope

This standard specifies the sugarcane stripe mosaic virus (Sugarcanestreakmosaicvirus) real-time fluorescent reverse transcription polymerase chain reaction Instruments and equipment, reagents and materials, sample collection and pre-processing, detection, and judgment and presentation of results should be performed (RT-PCR). This standard applies to the rapid detection and diagnosis of living host plants that may have sugarcane stripe mosaic virus.

2 Abbreviations

The following abbreviations apply to this document. CP. coat protein Ct value. the number of cycles experienced by the fluorescent signal in each reaction tube reaching the set threshold in a real-time fluorescent PCR reaction (cycle Threshold) cDNA. single-stranded DNA (complementaryDNA) complementary to the RNA strand ExTaqHS enzyme. hotstart DNA polymerase (hotstartDNApolymerase) FAM. 6-carboxy-fluorescein PCR. polymerase chain reaction (polymerase chain reaction) RNA. ribonucleic acid RT-PCR. reverse transcription polymerase chain reaction (reverse transcription polymerase chain reaction) SCSMV. Sugarcane Stripe Mosaic Virus (Sugarcanestreakmosaicvirus) TAMRA. 6-carboxytetramethylrhodamine Tip. Tip

3 Principle of the method

Design a pair of specific primers and a special one that are conserved only between the CP genes of the virus according to the CP gene sequence of sugarcane stripe mosaic virus A heterologous fluorescent double-labeled hydrolyzed oligonucleotide probe (TaqMan probe). First reverse the viral RNA with reverse transcriptase cDNA, and then real-time fluorescent PCR amplification reaction in the same reaction tube using cDNA as a template, hydrolyzed oligonucleotide probe The fluorescence signal intensity of the (TaqMan probe) is proportional to the increase of the PCR product of the target sequence, and is amplified by collecting PCR. By increasing the fluorescence signal value of the process, it is possible to determine whether the sample carries the target sequence. See Appendix A for information on sugarcane streak mosaic disease.

4 Instruments and equipment

4.1 Real-time fluorescent PCR detector. excitation/detection wavelength range 350nm~750nm; available fluorescent dye (SYBRGreenI) and water Decoded oligonucleotide probe (TaqMan probe); temperature rise and fall ≥2.0 °C/s; uniformity /-0.5 ° C; accuracy /-0.3 ° C; temperature The range is from 4 ° C to 100 ° C. 4.2 Nucleic acid protein analyzer or UV spectrophotometer. 4.3 Electronic balance. Sensitivity 0.01g. ......

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