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GB/T 28067-2011 English PDF

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GB/T 28067-2011: Detection of sugarcane yellow leaf virus using the real-time RT-PCR
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PDF similar to GB/T 28067-2011


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Basic data

Standard ID GB/T 28067-2011 (GB/T28067-2011)
Description (Translated English) Detection of sugarcane yellow leaf virus using the real-time RT-PCR
Sector / Industry National Standard (Recommended)
Classification of Chinese Standard B16
Classification of International Standard 65.020.01
Word Count Estimation 10,196
Date of Issue 2011-12-30
Date of Implementation 2012-06-01
Regulation (derived from) Announcement of Newly Approved National Standards No. 23 of 2011
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 sugarcane yellow leaf virus real-time fluorescent RT-PCR detection instruments and reagents required for sample collection and pre-treatment, methods of operation and the results determined. This standard applies to sugar cane plants, seedlings and seed stems of sugarcane yellow leaf virus for rapid detection, diagnosis.

GB/T 28067-2011: Detection of sugarcane yellow leaf virus using the real-time 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 of sugarcane yellow leaf virus using the real-time RT-PCR ICS 65.020.01 B16 National Standards of People's Republic of China Sugarcane yellow leaf virus real-time fluorescent RT-PCR detection method Issued on. 2011-12-30 2012-06-01 implementation Administration of Quality Supervision, Inspection and Quarantine of People's Republic of China Standardization Administration of China released

Foreword

This standard was drafted in accordance with GB/T 1.1-2009 given rules. This standard by the National Standardization Technical Committee of Plant Quarantine (SAC/TC271) and focal points. This standard was drafted. Sugarcane Research Institute, Fujian Agriculture and Forestry University, Ministry of Agriculture, sugar cane and products Quality Supervision and Testing Center, Agriculture Ministry Key Laboratory of Sugarcane Genetic Improvement. The main drafters of this standard. Three Bases, Chenping Hua, Chen Rukai, Guo Jinlong, Zhang Hua, Xu Liping, Wang Hengbo, Chen from strong. Sugarcane yellow leaf virus real-time fluorescent RT-PCR detection method

1 Scope

This standard specifies the collection instruments and reagents Sugarcane yellow leaf virus real-time fluorescent RT-PCR detection methods required for the sample with the front office Management, methods of operation and the results of determination. This standard applies to sugar cane plants, seedlings and seed stem rapid detection of sugarcane yellow leaf virus, diagnosis.

2 Terms and definitions

The following terms and definitions apply to this document. 2.1 Reverse transcription reversetranscription RNA as a template DNA synthesis process, also known as reverse transcriptase. 2.2 Real-time fluorescent RT-PCR realtimeRT-PCR Real-time fluorescence reverse transcription - polymerase chain reaction. 2.3 Ct values \u200b\u200bcycletime The number of cycles when fluorescence signal of each reaction tube reaches a set threshold experienced.

3 Abbreviations

The following abbreviations apply to this document. RNA. ribonucleic acid Taq enzyme. TaqDNA polymerase dNTPs. 4-deoxy-5'-triphosphate Zhong mixture RNase. RNA enzyme M-MLV. Moloney murine leukemia virus (Moloneymurineleukeminvirus) reverse transcriptase FAM. 6- carboxyfluorescein TAMRA. 6- carboxy-tetramethylrhodamine

4 principle of the method

In the reverse transcriptase enzyme RNA was reverse transcribed into cDNA, and then to take advantage of Taq DNA polymerase cDNA as a template for real-time fluorescence PCR amplification By reaction (using TaqMan probe method). On the basis of comparison of sugarcane yellow leaf virus coat protein gene on to design a pair just in sugarcane yellow Inter mosaic virus coat protein gene-specific primers and conserved a specific dual-labeled fluorescent probes. 5 'end of the probe labeled fluorescent FAM Light elements for the report fluorophore, the 3 'end is labeled TAMRA fluorescence quenching fluorophore binding site located inside the object amplified fragment. When the complete sequence of the probe and paired fluorophore fluorescence emitted by the 3 'end of the proximity of the quencher is quenched, the instrument can not be detected The fluorescence signal; however, when carrying out the extension reaction, Taq polymerase play a 5 '→ 3' exonuclease function of the degradation of the probe, such that the fluorophore

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