SN/T 5480-2022 in English

VALID

Technical specification for rainbow trout species identification-real-timePCR method

Issued on:2022-07-07
Implemented on:2023-02-01
File Format:PDF
Delivery:Via email within 1~3 business days

Price(USD)： $210.00
$204.00

Standard No:	SN/T 5480-2022
Document status:	VALID
Title in English:	Technical specification for rainbow trout species identification-real-timePCR method
Title in Chinese:	虹鳟成分鉴定技术规范实时荧光PCR法
Language:	English
File Format:	Electronic (PDF)
Delivery:	Via email within 1~3 business days
Issued on:	2022-07-07
Implemented on:	2023-02-01
Professional Classification:	SN-Import&Export Inspection
Related Keywords:	rainbow trout species rainbow trout standard technical framework technical specification identification-real-timepcr method introduction analysis
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Introduction

Analysis of the standard technical framework

This standard uses a real-time fluorescence PCR instrument to detect specific fragments of mitochondrial DNA and uses a 102bp amplification product to identify the ingredients of rainbow trout (Oncorhynchus mykiss) and Gill salmon. The core technology includes:

Technical elements	Parameter requirements	Quality control
Primer probe	OO.mykiss-F/R/P sequence	100pmol/L storage solution
Reaction system	12.5μL MasterMix	Double parallel control
Cycling conditions	95℃ 10min→40 cycles	Ct value judgment interval

Key implementation points

1. Sample processing specifications

Muscle tissue sampling should avoid fat contamination. Non-lethal sampling can use 3-5mm fin ray tissue. Three samples need to be packaged for testing/retesting/preserving during pretreatment, and 200mg solid or 200μL liquid is the standard sampling volume.

2. DNA extraction optimization

Protease K digestion combined with phenol/chloroform extraction is recommended, and the A260/A280 ratio is required to be between 1.7-2.0. Commercial kits need to pass equivalence verification.

3. Result interpretation rules

Positive judgment: Ct≤35.0 and typical amplification curve
Gray area processing: 35.0
Species confirmation: Need to combine SN/T 5203 gene barcode

Technology evolution analysis

Compared with traditional morphological identification, this standard has the following advantages:

Sensitivity advantage: Can detect <1% adulteration of ingredients in processed products

Timeliness breakthrough: Complete from DNA extraction to result interpretation within 6 hours

Degree of standardization: Realize comparability between laboratories through unified primer and probe sequences and Ct value threshold setting

Industry application suggestions

It is recommended to use this method in the following scenarios:

Authenticity verification of imported salmon products
Monitoring of germplasm resources in the breeding process
Traceability of aquatic products on e-commerce platforms

Pay attention to the regular calibration of the centrifuge and PCR instrument during implementation. It is recommended to add quality control samples to each batch of testing.

More samples: JC/T 2453-2018 DL/T 5154-2012 DL/T 368-2010

Sample only — not a preview of SN/T 5480-2022

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