SN/T 5334.6-2020 in English

VALID

Protocol of digital PCR for quantitatively detecting genetically modified plants and their derived products-Part 6: Genetically modified potato

Issued on:2020-12-30
Implemented on:2021-07-01
File Format:PDF
Delivery:Via email within 1~3 business days

Price(USD)： $150.00
$146.00

Standard No:	SN/T 5334.6-2020
Document status:	VALID
Title in English:	Protocol of digital PCR for quantitatively detecting genetically modified plants and their derived products-Part 6: Genetically modified potato
Title in Chinese:	转基因植物产品的数字PCR检测方法第6部分：转基因马铃薯
Language:	English
File Format:	Electronic (PDF)
Delivery:	Via email within 1~3 business days
Issued on:	2020-12-30
Implemented on:	2021-07-01
ICS Classification:	65.020.01-Farming and forestry in general
Chinese Classification:	B16-Plant quarantine, pest control
Professional Classification:	SN-Import&Export Inspection
Related Keywords:	digital pcr proven commercial digital pcr platform perform copies/μl key detection elements target gene design potato core technology innovation technical parameters traditional qpcr digital pcr potato av43 line
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本文件规定了进、出口马铃薯中转基因品系 AV43 特异性的数字 PCR（dPCR）定量检测方法。
本文件适用于马铃薯中转基因马铃薯 AV43 品系特异性的成分定量检测。本文件检测限参见附录 A。

Introduction

Technical background of the standard

This standard, as part 6 of the SN/T 5334 series, establishes a digital PCR quantitative detection system for the genetically modified potato AV43 line. As the global trade volume of genetically modified crops grows, the General Administration of Customs will implement this standard in 2021 to strengthen port quarantine capabilities.

Core Technology Innovation

Technical Parameters	Traditional qPCR	Digital PCR(dPCR)
Detection Principle	Fluorescence Signal Threshold Cycle	Absolute Droplet Counting
Quantification Method	Relative Quantification	Absolute Quantification
Limit of Detection (LOD)	1.5 copies/μL	1.2 copies/μL

Key Detection Elements

Target gene design

Potato β-sucrose invertase (fru) was used as the internal reference gene, the 5' end binding region of the exogenous sequence of the AV43 strain was used as the specific target, and the probes all used the FAM-BHQ1 fluorescent system.

Reaction system optimization

The 25μL system contained 12.5μL 2×ddPCR Super Mix, the final concentration of primers/probes was 200nmol/L, and the DNA template loading amount was 4ng/μL.

Implementation recommendations

Give priority to a proven commercial digital PCR platform
Perform the amplification procedure strictly in accordance with the annealing temperature (60°C) in Appendix A
A positive result must satisfy both the internal reference gene Ct≤35 and the exogenous gene copy number ≥3

More samples: JJG 21-2008 GB 18306-2015 GB 50021-2001(2009)

Sample only — not a preview of SN/T 5334.6-2020

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