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GB/T 36829-2018: Detection of Leifsonia xyli subsp. xyli by real-time PCR
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Detection of Leifsonia xyli subsp. xyli by real-time PCR
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GB/T 36829-2018
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Basic data | Standard ID | GB/T 36829-2018 (GB/T36829-2018) | | Description (Translated English) | Detection of Leifsonia xyli subsp. xyli by real-time PCR | | Sector / Industry | National Standard (Recommended) | | Classification of Chinese Standard | B16 | | Classification of International Standard | 65.020.01 | | Word Count Estimation | 10,190 | | 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 36829-2018: Detection of Leifsonia xyli subsp. xyli by real-time 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 of Leifsonia xyli subsp.xyli by real-time PCR
ICS 65.020.01
B16
National Standards of People's Republic of China
Sugarcane
Real-time fluorescent 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, Sun Shengren, Wang Hengbo, Fu Huaying, Huang Meiting, Wang Jinda, Zhang Huili, Chen Rukai.
Introduction
The issuing authority of this document draws attention to the fact that when the statement is in compliance with this document, it may involve real-time fluorescence quantification with sugarcane rat root stunting bacteria.
Utilization of PCR related patents.
The issuing organization of this document has no position on the authenticity, validity and scope of the patent.
The holder of the patent has assured the issuing authority of this document that he is willing to work with any applicant on reasonable and non-discriminatory terms and conditions.
Negotiate a patent license. The patent holder's statement has been filed with the issuing authority of this document. Related information can be passed the following
Contact information obtained.
Patent holder name. Wang Hengbo.
Address. No. 15, Shangxia Road, Cangshan District, Fuzhou City, Fujian Province.
Please note that in addition to the above patents, certain aspects of this document may still involve patents. The issuing organization of this document does not undertake to identify these special
Liability.
Sugarcane
Real-time fluorescent PCR detection method
1 Scope
This standard specifies the instrument for real-time fluorescent PCR detection of sugarcane rat root stunting disease (Leifsoniaxylisubsp.xyli)
Equipment, reagents and materials, sample collection and pre-treatment, detection and results judgment and presentation.
This standard is applicable to the rapid detection and diagnosis of living host plants that may have cane root stunting disease.
2 Abbreviations
The following abbreviations apply to this document.
CTAB. CetyltrimethylAmmonium Bromide
Ct value. Cycle Threshold (CycleThreshold)
DNase. Deoxyribonuclease
FAM. 6-Carboxyfluorescein (6-Carboxy-Fluorescein)
Lxx. sugarcane rat root stunting disease (Leifsoniaxylisubsp.xyli)
Part-1. a pathogenic gene encoded by the genome of sugarcane rat root stunting disease (Pathogenicity Gene-1)
PCR. Polymerase Chain Reaction (PolymeraseChainReaction)
RNase. Ribonuclease
TAMRA. 6-Carboxytetramethylrhodamine
3 Basic information on sugarcane ratoon stunting bacteria
The sugarcane rat root stunting disease belongs to the member of the Leifsonia family of the genus Leifsonia of the Microbacteriaceae family.
The name is Leifsoniaxylisubsp.xyli, abbreviated as Lxx. See Appendix A for additional information on sugarcane rat root stunting bacteria.
4 Principle of the method
Designing a pair of specific primers that are conserved only between the Pat-1 gene of the pathogen based on the pathogenic gene Pat-1 sequence of the stunting dwarf pathogen
A specific fluorescent double-labeled TaqMan probe. Fluorescence signal intensity of TaqMan probe in real-time fluorescent PCR amplification
Along with the increase of the PCR product of the target sequence, the fluorescence signal value of the PCR amplification process can be judged.
Whether the sample has a target sequence.
5 Instruments and equipment
5.1 Real-time fluorescent PCR detector. excitation/detection wavelength range 350nm~750nm; SYBRGreenI dye and
TaqMan probe; temperature rise and fall ≥2.0 °C/s; uniformity ± 0.5 ° C; accuracy ± 0.3 ° C; temperature range 4 ° C ~ 100 ° C.
5.2 Nucleic acid protein analyzer or UV spectrophotometer.
5.3 Electronic balance. Sensitivity 0.01g.
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