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Extraction method and applications of inset dry specimen genome

An extraction method and dry specimen technology, applied in the field of molecular biology, can solve problems such as hindering the research process of molecular phylogenetic evolution of insects, the inability to use genetic gene information, and the inability of insects to quickly and effectively determine the extraction method, achieving simple and easy operation. The effect of high efficiency, high extraction efficiency and wide application range

Inactive Publication Date: 2014-01-15
NORTHWEST A & F UNIV
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Problems solved by technology

However, due to the large number of insect species, the huge system, and the differences in developmental stages, the extraction methods of insects not involved in the research cannot be quickly and effectively determined, so that the huge genetic information library contained in the insect herbarium cannot be used. Hindered progress in the study of molecular phylogenetic evolution in insects

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  • Extraction method and applications of inset dry specimen genome
  • Extraction method and applications of inset dry specimen genome
  • Extraction method and applications of inset dry specimen genome

Examples

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Embodiment 1

[0048] Example 1 Improved Saturated Sodium Chloride Method to Extract Genomic DNA from Dried Specimens of Different Orders of Insects and Its Detection

[0049] 1. DNA extraction

[0050] (1) Obtain 0.03 g of the sample used to extract genomic DNA from the head, thorax, and foot of the insect dry specimen, in STE buffer (0.1mmol / L NaCl, pH8.0 10mmol / L Tris buffer, pH8.0 1mmol / LEDTA, dsH2O) at room temperature for 5 hours, and then dried on sterilized filter paper.

[0051] (2) Put the dry pretreatment material into the Ep tube and grind it to powder with a sterilized glass rod, transfer the ground product into the Ep tube,

[0052] (3) Add 350 μl of solution 1 (0.4 ml TE containing 0.1% RNase) to dissolve and remove RNA;

[0053] (4) Add digestion solution 2 including 2-8% SDS, 100-300 μg / ml proteinase K, pH 8.0) digestion temperature is 25-37°C for 10 minutes, centrifuge, transfer the supernatant to another 1.5ml Ep tube,

[0054] (5) Protein precipitation:

[0055] 1) A...

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Abstract

The invention discloses an extraction method and applications of inset dry specimen genome DNA. The extraction method is based on improvement of saturated NaCl method, and comprises following steps: (1) pretreatment of insect dry specimen; (2) the pretreated material is grinded into powder, and the powder is delivered into a 1.5ml EP tube; (3) solution 1 is added for dissolving so as to remove RNA; (4) digestion liquid 2 is added, the solution is centrifuged, and supernatant is delivered into another EP tube; (5) protein is precipitated; and (6) washing. The extraction method is suitable for DNA extraction of different parts of a plurality of kinds of insect dry specimens such as hymenoptera, odonata, orthoptera and coleoptera; and is high in extraction efficiency. Quality and purity of extracted DNA are capable of meeting requirements of insect molecule system evolution analysis. Basic methods for research and exploration on extraction of insect dry specimen genome DNA at the most appropriate parts with least material amount and least damages are provided. The extraction method is simple and convenient for operation, and applications of the extraction method in labs are convenient.

Description

technical field [0001] The invention relates to the technical field of molecular biology, in particular to a method for extracting genome DNA of insect specimens. Background technique [0002] DNA is the carrier of genetic information and the main object of molecular biology research. Since the DNA sequence contains rich biological information, it records the history of biological evolution. In living cells, there are intact DNA repair mechanisms that keep DNA intact. However, the chemical properties of DNA are extremely unstable and can be degraded spontaneously through hydrolysis or oxidation in dry samples. The dry specimen in the herbarium is a huge genetic information library, modern molecular biology random amplified polymorphic DNA (RAPD) technology, restriction fragment length polymorphism (RFLP) technology DNA fingerprint technology, and 12srRNA, The determination of 16srRNA, 18SrRNA, ND5 and other sequences has been widely used in the study of insect classificat...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/10
Inventor 南小宁伏正魏琮冯继广周浪赵雅琼郭新荣
Owner NORTHWEST A & F UNIV
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