SN/T 5334.2-2020 in English

VALID

Protocol of digital PCR for quantitatively detecting genetically modified plants and their derived products - Part 2: Genetically modified soybean

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

Price(USD)： $160.00
$156.00

Standard No:	SN/T 5334.2-2020
Document status:	VALID
Title in English:	Protocol of digital PCR for quantitatively detecting genetically modified plants and their derived products - Part 2: Genetically modified soybean
Title in Chinese:	转基因植物产品的数字PCR检测方法第2部分：转基因大豆
Language:	English
File Format:	Electronic (PDF)
Delivery:	Via email within 1~3 business days
Issued on:	2020-12-30
Implemented on:	2021-07-01
Professional Classification:	SN-Import&Export Inspection
Related Keywords:	digital pcr quantitative pcr detection digital pcr real-time fluorescence conventional pcr detection technology droplet pcr
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Introduction

Analysis of the standard technical framework

Core elements	Requirements of SN/T 5334.2	Comparison with conventional PCR
Detection technology	Droplet digital PCR	Real-time fluorescence quantitative PCR
Detection objects	13 genetically modified strains including FG72	Usually single strain detection
Quantitative limit	1.0-3.0 copies/μL	10-100 copies/μL

Key Technical Innovations

The standard adopts the dual probe labeling strategy (FAM/BHQ1 and TAMRA), which is solved through cross-border sequence design:

3'-end specific detection of DAS68416-4 strain
5'-end junction sequence capture of MON87708 strain

The cycling parameters are set differently according to the GC content, such as the CV127 strain requires a denaturation time of 30s.

Implementation and Application Guide

Typical Detection Process

Sample preparation: soybean seed grinding particle size ≤ 0.5mm
DNA extraction: CTAB method purity requirement A260/280=1.8-2.0
Reaction system: 60ng template DNA+2×ddPCR Super Mix
Thermal cycle program: 94℃ denaturation 15-30s, 60℃ annealing 44-49 cycles

Key points of quality control

Negative control: non-GMO soybean DNA
Positive control: standard substances of various strains (ERM-AD413, etc.)
Blank control: ultrapure water instead of template

Background of Standard Evolution

With the commercial planting of new insect-resistant soybeans such as DAS-81419-2 in Brazil in 2019, the traditional qPCR method has shown limitations in:

Multiple target detection
Low-content sample quantification
Complex matrix anti-interference

, which prompted the formulation of this standard.

More samples: NB/T 47013.2-2015 DL/T 368-2010 DL/T 5154-2012

Sample only — not a preview of SN/T 5334.2-2020

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