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SN/T 2497.22-2010: Test method of import and export dangerous chemicals. Part 22: Determination of DNA Tm
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Test method of import and export dangerous chemicals. Part 22: Determination of DNA Tm
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SN/T 2497.22-2010
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Basic data | Standard ID | SN/T 2497.22-2010 (SN/T2497.22-2010) | | Description (Translated English) | Test method of import and export dangerous chemicals. Part 22: Determination of DNA Tm | | Sector / Industry | Commodity Inspection Standard (Recommended) | | Classification of Chinese Standard | G09 | | Classification of International Standard | 13.300 | | Word Count Estimation | 4,422 | | Date of Issue | 2010-03-02 | | Date of Implementation | 2010-09-16 | | Regulation (derived from) | National Quality Inspection (2010) 98; industry standard filing Notice 2010 No. 5 (No. 125 overall) | | Issuing agency(ies) | General Administration of Customs | | Summary | This standard specifies the import and export of dangerous chemicals Tm value of DNA test sample measured, solutions and test procedures. This standard applies to DNA tests import and export of dangerous chemicals Tm values ??measured. | SN/T 2497.22-2010: Test method of import and export dangerous chemicals. Part 22: Determination of DNA Tm
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Test method of import and export dangerous chemicals.Part 22. Determination of DNA Tm
Exit inspection and quarantine industry standard book People's Republic of China
Import and export of dangerous chemicals - Test methods
Issued on. 2010-03-02
2010-09-16 implementation
People's Republic of China
The State Administration of Quality Supervision, Inspection and Quarantine released
Foreword
SN/T 2497 "Test Methods for Chemical Safety & Exports dangerous" series of standards is divided into 29 parts.
--- Part 1. mammalian liver cells in vivo DNA synthesis (UDS) test;
--- Part 2. plaque-forming cells (PFC) test;
--- Part 3. Large Zhuan reproduction test;
--- Part 4. Saccharomyces cerevisiae mitotic recombination test;
--- Part 5. testicular cells UDS test;
--- Part 6. mammalian cells in vitro sister chromatid exchange test;
--- Part 7. mouse ear swelling test;
--- Part 8. popliteal lymph node assay;
--- Part 9. Determination of serum hemolysin test;
Part --- Article 10. T lymphocyte proliferation assay test;
--- Part 11. germline mutation assay;
--- Part 12. single cell gel electrophoresis test;
--- Part 13. fluorescence in situ hybridization;
Part --- Article 14. SDS- polyacrylamide gel electrophoresis;
--- Part 15. PCR-SSCP experiment;
Part --- Article 16. Western-Blot test;
--- Part 17. Behavior of mammalian toxicology tests;
--- Part 18. Detection of DNA adducts;
--- Part 19. NorthernBlot experiment;
--- Part 20. Bradford protein assay;
Part --- Zi 21. agarose gel electrophoresis;
--- Part 22. Tm value determination method of DNA;
--- Part 23. organelle separation experimental methods;
--- Part 24. cellular immunity in vitro assay method;
--- Part 25. humoral immune function tests;
--- Part 26. macrophage function tests;
--- Part 27. Flow cytometry of apoptosis;
--- Part 28. Application shuttle plasmid mutations Inspection;
--- Part 29. Biochemical Oxygen Demand (BOD) was measured.
This part is Part 22 SN/T 2497 series of standards.
This section proposed and managed by the National Certification and Accreditation Administration Committee.
This section is responsible for drafting unit. People's Republic of China Tianjin Entry-Exit-Entry Inspection and Hunan People's Republic of China
Quarantine Bureau.
The main drafters of this section. Jia Xiaochuan, Wangli Bing, Lu Gang, Li Jing, Li Xue Yang, Chen Chun.
The first part of the Department of Inspection and Quarantine issued by industry standards.
Import and export of dangerous chemicals - Test methods
1 Scope
SN/T 2497 provisions of this part of the test sample and export of dangerous chemicals in the test Test Tm value of DNA,
Solution, test procedures and test results.
This section applies to the import and export of dangerous chemicals Tm value measurement test DNA.
Principle 2
DNA in when the temperature rose to a certain value, the hydrogen bonds between the two single-stranded nucleotide began to be opened (DNA begin to denature)
Separation, so that the DNA solution is significantly increased absorption in the ultraviolet at a wavelength of 260nm; When the temperature up to a certain value, complete separation of DNA
Into single strands, then continue to heat, UV absorption of DNA solution is no longer increases. When DNA thermal denaturation UV absorption maximum increase
Thermal denaturation temperature or melting temperature of the temperature corresponding to half of the DNA is called, i.e. the Tm value.
3 Reagents
3.1 Water
This standard test method involves the water are sterile distilled water.
3.2 Sodium chloride - sodium citrate buffer (SSC)
Sodium chloride and trisodium citrate were dissolved in a certain amount of water, such that the sodium chloride concentration of 0.15M solution, the concentration of sodium citrate
0.015M.
4 Test Procedure
4.1 DNA concentration
UV spectrophotometer, determination of DNA sample at 260nm absorbance value A, A × 50μg/mL, the concentration of DNA was calculated.
Sample preparation 4.2
DNA samples taken 60μg, diluted with 3mL SSC to (0.3 to 0.4 absorbance).
4.3 Sample purity
260nm, 280nm and 230nm absorption at the time by UV spectrophotometer to SSC as a reference solution, the samples were measured 25 ℃
Photometric values A, when 1.75 <
A260
A280 < 1.85
And 0.5 <
A230
A260 < 0.6
When the determination indicates that the sample purity requirements, you can Tm value
Determination.
4.4 Determination
With the temperature programmed by means of UV spectrophotometer to SSC as a reference solution, the sample was measured by the absorbance at 25 ℃ ~ 99 ℃ of A
(Later increased by 75 ℃ 1 ℃, a record absorbance value; generally 25 ℃ ~ 75 ℃, the absorbance changes little; no absorbance after 94 ℃
Variety).
5 results
Shall absorbance multiplied by the relative swelling volume when the temperature record and the corresponding absorbance plotted DNA thermal denaturation curve (accurate measurement
Vp T/Vp 25, corrected for the absorbance of an aqueous solution of 25 volume deg.] C), the thermal denaturation midpoint temperature curve corresponding to the Tm value.
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