小蠹科昆虫基因组DNA提取方法的比较研究
Comparison of four genomic DNA extraction methods with Scolytidae beetles.
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摘要: 为探讨小蠹科昆虫基因组DNA的提取方法，分别以单头削尾材小蠹、削尾材小蠹足部肌肉组织以及多年75%酒精浸渍标本为研究对象，采用经典的酚仿抽提法、改良的CTAB法、SDS法及动物组织/细胞基因组DNA提取试剂盒(即磁珠法)4种方法提取小蠹基因组DNA，对提取的基因组DNA进行分光光度值的测定以及COⅠ基因扩增。结果表明：4种方法均可从单头标本中提取总DNA；对于75%酒精浸渍2年的标本，4种方法均适用，大于2年的浸渍标本，仅磁珠法适用，且磁珠法适合微量组织总DNA的提取。Abstract: Four methods used for extracting the genomic DNA of bark beetles, Xyleborus mutilatus (Coleoptera, Scolytidae) were studied in this article. An individual X. mutilatus beetle, a bark beetle leg and the specimen reserved in 75% ethanol were selected as testing sample respectively, the genomic DNA of bark beetle was extracted by using four different methods, including classical phenol/chloroform extracting method, improved cetyltrimethyl ammonium bromide (CTAB) method, the sodium dodecyl sulfate (SDS) method and animal tissue/cell genomic DNA extraction kit (i.e. paramagnetic particle method). The extracted genomic DNA was treated with photometric analysis and COⅠ gene amplification. The results showed that all the four methods can be used for the total DNA extraction. For the specimen soaked in 75% ethanol, these four methods were all applicable, however, only the paramagnetic particle method was suitable for the DNA extraction of the specimen,which had been soaked in 75% ethanol over two years. Furthermore, the paramagnetic particle method was also applicable for the total DNA extraction from microscale tissue. The study provids primary information for further molecular systematic research of bark beetles.
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Keywords:
- Scolytidae /
- genomic DNA /
- COⅠ /
- molecular identification
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