Traditional Chinese medicinal material genome DNA extraction method suitable for DNA amplification technology

An extraction method and a technology of traditional Chinese medicinal materials, which are applied in the field of molecular biology, can solve problems such as cumbersome procedures, damage to the health of operators, and complicated operations, and achieve the effects of strong versatility, saving reagents and consumables, and little pollution

Inactive Publication Date: 2014-03-12
INST OF CHINESE MATERIA MEDICA CHINA ACAD OF CHINESE MEDICAL SCI
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  • Abstract
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  • Application Information

AI Technical Summary

Problems solved by technology

However, the CTAB method or SDS method is cumbersome and complicated to operate, and generally takes more than 5 hours to extract DNA
And a large amount of organic solvents such as phenol / chloroform, β-mercaptoethanol, and isopropanol are used in the operation process, which is detrimental to the health of operators

Method used

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  • Traditional Chinese medicinal material genome DNA extraction method suitable for DNA amplification technology
  • Traditional Chinese medicinal material genome DNA extraction method suitable for DNA amplification technology
  • Traditional Chinese medicinal material genome DNA extraction method suitable for DNA amplification technology

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Experimental program
Comparison scheme
Effect test

specific Embodiment 2

[0033] 1. Materials

[0034] The materials are black snake, money snake, earth dragon, and scorpion; the method is the same as the implementation case 1, and the difference between this implementation case and the implementation case 1 is the use of the black snake specific identification primer WSS-1: 5'-GCGAAAGCTCGACCTAGCAAGGGGACCACA- 3', and WSS-2: 5'-CAGGCTCCTCTAGGTTGTTATGGGGTACCG-3'; PCR reaction system: 25 μL PCR reaction system containing 2.5 μL 10×ExTaq buffer, 2 μL 0.25 mmol / L dNTPs; 0.25 pmol each of upstream primer and downstream primer; 0.8 U Ex taq DNA polymerase, 1 μL of the above DNA template. The reaction was amplified on the 9700 gene amplification instrument of ABI company, and the cycle parameters were pre-denaturation at 94°C for 5 min; denaturation at 94°C for 30 s, extension at 63°C for 30 s, 38 cycles; extension at 72°C for 7 min; storage at 4°C after the reaction was completed. .

[0035] 2. Results

[0036] According to the method used in the presen...

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Abstract

The invention discloses a genome DNA rapid extraction method suitable for the DNA amplification technology. The extraction method is quite simple and economic as only sodium hydroxide solution and Tris-HCl solution are employed. By employing the extraction method, DNA of a sample can be totally extracted just by 15 minutes. The extraction method is universal, and good extraction effects on decocting pieces of plant medicine and animal medicine, and processed products of the decocting pieces can be achieved. The quality of the extracted DNA meets demands of downstream PCR amplification, problems of long DNA extraction time and heavy workload in a traditional Chinese medicinal material common PCR reaction template are solved, and the method meets demands of DNA being rapidly extracted in scientific researches and production processes.

Description

technical field [0001] The invention relates to a method for rapidly extracting genomic DNA of Chinese medicinal materials, belonging to the technical field of molecular biology. Background technique [0002] The PCR reaction (polymerase chain reaction), which came out in 1985, is an in vitro nucleic acid amplification technology. It has been widely used because of its high sensitivity, strong specificity, simple operation, good repeatability, and easy automation. Various fields of life sciences, medicine, and pharmacy have become indispensable means of medical research. In addition, isothermal amplification techniques include Q replicase reaction (QBRA), strand displacement amplification (SDA), nicking enzyme constant temperature amplification (NEMA), transcription-dependent amplification (TASTMA, 3SR, NASBA) , helicase amplification (HAD), rolling circle amplification (RCA), loop-mediated isothermal amplification (LAMP), single primer isothermal amplification (SPIA), etc....

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/10
Inventor 黄璐琦袁媛蒋超
Owner INST OF CHINESE MATERIA MEDICA CHINA ACAD OF CHINESE MEDICAL SCI
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