SN/T 5200-2020 in English
VALIDProtocol for species identification of Manis Spp.
- Issued on:2020-12-30
- Implemented on:2020-07-01
- File Format:PDF
- Delivery:Via email within 1~3 business days
$262.00
| Standard No: | SN/T 5200-2020 |
| Document status: | VALID |
| Title in English: | Protocol for species identification of Manis Spp. |
| Title in Chinese: | 穿山甲物种鉴定技术规范 |
| Language: | English |
| File Format: | Electronic (PDF) |
| Delivery: | Via email within 1~3 business days |
| Issued on: | 2020-12-30 |
| Implemented on: | 2020-07-01 |
| Professional Classification: | SN-Import&Export Inspection |
| Related Keywords: | species identification
improvement points species new species core detection technology morphological identification system species |
| Related Topics: | species
species identification species identification Several mass spectrometric identification techniques Rubber identification Document Production Time Identification Technical Specification Identification Identification of four substances QB/T 5200 Animal breed identification industry Polymer type identification Specifications for identification of fishery germplasm Pangolin species identification technology |
Introduction
Analysis of the Standard Technical Framework
| Technical Dimensions | SN/T 5200-2020 | SN/T 1162-2002 | Improvement Points |
|---|---|---|---|
| Species Covered | 7 species of pangolins | 4 species | 3 new species including tree pangolins |
| Detection Methods | SYBR Green I Real-time Fluorescence PCR | Conventional PCR | Sensitivity increased by 100 times |
| Judgment Criteria | Ct value + Tm value + sequencing | Electrophoresis bands | Triple verification mechanism |
Detailed explanation of core detection technology
Morphological identification system
The standard provides a matrix of identification characteristics for seven species, including Chinese pangolin and Malay pangolin:
- Scale structure: Chinese pangolin scales are fan-shaped (560-688 pieces), and Malayan pangolins have sharp ridge scales
- Body size ratio: The tail length/body length ratio of tree pangolins is 1.25:1, while that of South African pangolins is only 0.85:1
- Special marks: The anal glands of Indian pangolins secrete yellow defensive liquid
Optimization of molecular detection process
Amplification of 206bp fragment of Cytb gene using MAF/MAR primer pair:
- DNA extraction: TES buffer + SDS lysis, phenol-chloroform purification
- Reaction system: 20μL containing SYBR Premix Ex Taq enzyme
- Amplification program: 95℃ 2min initial denaturation, 40 cycles (95℃ 15s→60℃ 30s)
Interpretation threshold: Ct<35 and Tm value match is positive, 35≤Ct<40 needs to be retested for confirmation
Standard implementation recommendations
Laboratory configuration requirements
| Equipment type | Technical parameters | Calibration requirements |
|---|---|---|
| Real-time fluorescence PCR instrument | Melting curve analysis function | Quarterly calibration |
| Electrophoresis system | Voltage range 50-150V | Monthly verification |
Key points of quality control
- Each batch of experiments must include a positive control (pangolin genomic DNA) and a blank control
- Scale samples need to be surface decontaminated (washed with 0.85% saline)
- Sequencing results should be compared with the reference sequence in Appendix E for a ≥98% similarity
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