SN/T 3402-2012 in English

VALID

Identification genuineness and purity of two-line rice varieties using DNA analysis method

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Standard No:	SN/T 3402-2012
Document status:	VALID
Title in English:	Identification genuineness and purity of two-line rice varieties using DNA analysis method
Title in Chinese:	两系水稻品种真实性与纯度鉴定 DNA分析法
Language:	English
File Format:	Electronic (PDF)
Delivery:	Via email within 1~3 business days
Professional Classification:	SN-Import&Export Inspection
Related Keywords:	dna analysis method introduction technical background core detection principle detection target technical method judgment basis identification two-line rice varieties two-line hybrid rice dna concentration
Related Topics:	dna purity authenticity dna fragment analysis Two Methods of Thermal Analyzers Purity standard sn t 3372 2012 Vacuum analysis dna identification analysis Sample purity and content sn/t+3438-2012 sn/t+3372-2012 sn+t+3372+2012 dsc two methods sn/t+3367-2012 dna standard sn/t3367-2012 sn/t3209-2012 sn/t3372-2012 dna fragment analysis dna fragment analysis modern dna analysis dna molecular analysis forensic dna analysis Purity Analysis dna purifier Identification analysis dna/ dna analysis technology Analyze two lines Sample purity Qualitative analysis Two Methods of Purity Analyzers dna quantitative analyzer Peak Purity Moisture dna purity rice variety rice variety Four Quantitative Analysis Methods product dna analysis Differential Thermal Analysis Two dna analysis Identification analysis dna analysis dna strand analysis Analytical purity dna strand analysis Two functions of the product Two ways to measure cod Two implementations of - Unlike the previous two samples dna purity Sample purity Analysis of dna purity Rice line trace analysis of pure water dna Measurement and Analysis abundance dna dna qualitative method sn/t3395-2012 Two ways to assess accuracy Two methods of mass spectrometric identification Seven Methods of Qualitative Analysis To identify an analytical method Experimental moisture how to identify dna sample purity Specific rotation and purity Two xrf methods dna pcr analysis Identification and analysis of food Identification of moisture Qualitative Analysis and Quantitative Analysis of Water Quality How to Identify Rice Varieties dna flow analysis Identification of rice rice DNA Two chiral crystallization separation methods Enzyme purification and identification Analysis of DNA enzyme digestion experiment results The latest standards for authenticity identification of wheat varieties Purity Quantitative Analysis Method There are two methods of analyzing phase equilibrium. Two pharmaceutical analysis methods are equivalent Variety identification industry

Introduction

Technical background of the standard

This standard aims at the unique photothermosensitive nuclear male sterility characteristics of two-line hybrid rice, and establishes a DNA molecular identification system based on PCR amplification instrument and electrophoresis detection system. By detecting characteristic marker genes such as pms1/pms3/tms5 and combining 48 SSR loci analysis, the authenticity and purity of the variety can be double verified.

Core Detection Principle

Detection Target	Technical Method	Judgment Basis
Identification of Sterile Lines	Characteristic Marker PCR+Enzyme Digestion	pms1-S1(237bp)、pms3-Cl(931bp)、tms5-S2(387bp)
Variety Authenticity	SSR Polymorphism Analysis	48 Loci with Differences ≥2 Were Determined to Be Different Varieties
Hybrid Purity	Parental Complementary Band Type Detection	Calculation formula: P=(N_T-N_P)/N_T×100%

Key technical parameters

DNA extraction requirements: DNA concentration extracted by CTAB method ≥50ng/μL, A260/A280 ratio 1.8-2.0

PCR system: 20μL reaction volume containing 1×Buffer, 2.5mM Mg²⁺, 0.25mM dNTPs, 1U Taq enzyme

Electrophoresis conditions:

Agarose gel: 1.5% concentration, 120V electrophoresis for 30min
Polyacrylamide gel: 6% denaturing gel, 200V electrophoresis for 2h

Implementation points

Sample requirements: The test sample must contain at least 100 seeds or individual plants, with a storage period of ≥6 months
Control setting: Standard samples must be used as controls, and hybrids must test both parents at the same time
Primer verification: Polymorphism verification is required when the SSR primers in Appendix B are used for the first time
Silver staining notes: The development time should be controlled within 5-10 minutes to avoid too dark background

Comparison of technological evolution

Technical indicators	Traditional morphology	This DNA standard
Detection cycle	≥1 growing season	3-5 working days
Sample quantity	More than 1000 seeds	100 seeds
Environmental interference	Significant	None
Resolution	Variety level	Genotype level

More samples: QC/T 942-2021 DL/T 5154-2012 GB 50021-2001(2009)

Sample only — not a preview of SN/T 3402-2012

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