SN/T 5227.6-2019 in English
VALIDRapid detection of water buffalo derived ingredient in food for export-Recombinase- aid amplification (RAA) method
- Issued on:2019-12-27
- Implemented on:2020-07-01
- File Format:PDF
- Delivery:Via email within 1~3 business days
$127.00
Introduction
Technical background and evolution of standards
As the sixth part of the SN/T 5227 series of standards, this standard applies recombinase-mediated amplification technology (RAA) to the detection of buffalo-derived components for the first time. Compared with traditional PCR technology, it has technical advantages such as constant temperature amplification (37-42℃) and short reaction time (20min). This standard complements ISO 20836:2021 in technology, filling the gap in my country's standards in the field of rapid molecular detection.
Core Detection Principle
| Technical Elements | Traditional PCR | RAA Technology | Technical Advantages |
|---|---|---|---|
| Reaction Temperature | 94-60℃ Cycle | Constant Temperature 39℃ | No Thermal Cycler Required |
| Key Enzyme | Taq DNA Polymerase | SC-recA/BS-recA Recombinase + Bsu Polymerase | Strand Replacement Efficiency Increased by 3 Times |
| Detection Time | ≥90min | ≤20min | Efficiency Increased by 75% |
| Sensitivity | 0.1% | 0.1% | Equal sensitivity |
Analysis of key operation points
1. DNA extraction quality control
When using CTAB method for extraction, please note that: A260/A280 ratio 1.7-1.9 is the qualified range. When testing high-fat samples, it is recommended to increase the number of phenol-chloroform extractions to 2-3 times.
2. Real-time fluorescence RAA system configuration
Standard requirements for key component concentrations: recombinase protein ≥ 300ng/μL, SSB protein 225ng/μL, magnesium acetate buffer 280mol/L. In actual operation, it should be noted that dNTPs need to be stored away from light to prevent oxidative degradation.
Implementation suggestions and troubleshooting
False positive prevention:The laboratory needs to be strictly zoned and managed, and it is recommended to use the UNG enzyme treatment system. According to the verification data of Zhengzhou Customs in 2020, adding 1U UNG enzyme can reduce the false positive rate by 82%.
Critical value treatment:When the Ct value is 35-40 or the T value is 15-20min, it is recommended to: ①Recheck the quality of DNA extraction ②Change the reagent batch ③Add internal reference gene detection.
Loading PDF document...
Error loading PDF. Please make sure the file is valid and try again.
We also recommend
-
SN 0283-1994 in English
\"Method for Inspection of Dichlorodipyridinol Residues in Exported Poultry Meat\"
1994-02-18 -
SN/T 0244-1994 in English
\"General Rules for Inspection and Certification of Imported Mechanical and Electrical Instruments\"
1993-08-01 -
SN 184-1993 in English
\"Test Methods for Listeria Monocytogenes in Exported Foods\"
1993-10-08 -
SN/T 0420-2010 in English
Inspection method of Trichinella spiralis in pork for export—Magnetic stirrer method for pooled sample digestion
2010-11-01 -
SN/T 1422-2015 in English
Control standards for medical vector in entry-exit airplane
2015-12-04 -
SN/T 0440.1-1995 in English
Rules for the inspection of technical performance of househeld electric washing machines for export
1995-12-25
