SN/T 5334.8-2020 in English

VALID

Protocol of digital PCR for quantitatively detecting genetically modified plants and their derived products-Part 8: Genetically modified sugarbeet

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 5334.8-2020
Document status:	VALID
Title in English:	Protocol of digital PCR for quantitatively detecting genetically modified plants and their derived products-Part 8: Genetically modified sugarbeet
Title in Chinese:	转基因植物产品的数字PCR检测方法第8部分：转基因甜菜
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:	mainstream digital pcr platforms key points digital pcr simultaneous detection design reaction system standard detection elements internal standard gene probe
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本文件规定了进、出口甜菜中转基因品系特异性的数字 PCR（dPCR）定量检测方法。
本文件适用于甜菜中转基因 H7-1 品系特异性的成分定量检测。本文件的定量检测低限为 2.8 拷贝 /μL，定性检测低限为 0.56 拷贝 /μL。

Introduction

Analysis of the core content of the standard

Detection elements	Technical parameters	Innovation points
Quantitative detection limit	2.8 copies/μL	5 times the sensitivity of traditional qPCR
Primer design	GS gene (118bp) + H7-1 strain (107bp)	Dual-channel simultaneous detection design
Reaction system	20μL final system	Compatible with mainstream digital PCR platforms

Key points for technical implementation

Key control points:The experiment must strictly follow the reaction system configuration shown in Table 2, where the final concentrations of the internal standard gene probe (VIC labeled) and the H7-1 strain probe (FAM labeled) must be controlled at 0.2μmol/L.

Amplification procedure:A two-step amplification method (94℃ 15s→60℃ 1min) was used, with 49 cycles. The hot start step needs to be adjusted according to the instructions for different platforms.

Standard Application Cases

When a customs laboratory tested imported beet powder, it found that the copy number of the internal standard GS gene was 35.2 copies/μL, but the H7-1 strain was not detected. Finally, it issued a report that "no genetically modified beet H7-1 strain components were detected", which met the judgment requirements of Chapter 7 of the standard.

Technology Evolution Analysis

Compared with the 2015 version of the qPCR method: this standard has the following advantages: the detection limit is increased from 10 copies/μL to 2.8 copies/μL; the sugar beet genome boundary sequence is clarified for the first time (Appendix A); the absolute quantitative advantage of digital PCR is introduced to avoid the error caused by the standard curve.

More samples: DL/T 5154-2012 YD/T 3934-2021 GB 18306-2015

Sample only — not a preview of SN/T 5334.8-2020

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