叩甲科昆虫微量基因组DNA抽提方法的研究

摘要

叩甲科昆虫种类数量众多,分布广泛,很多种类都是重要的地下害虫.由于叩甲科昆虫外部形态相似度非常高,传统的形态分类工作具有较大的难度,对于叩甲科的系统发育研究大多基于分子标记进行展开.目前很多昆虫基因组的抽提方法会破坏整个虫体,为此,本研究改进了1种微量昆虫基因组DNA的抽提方法,采用蛋白酶K消化法,以叩甲昆虫的1条后足肌肉作为抽提对象,以期在不破坏叩甲主要鉴定特征的前提下,提取到较高质量的基因组DNA.同时,对提取到的基因组进行PCR扩增和测序,结果表明该方法抽提到的基因组DNA可以应用于分子学相关试验.由于该方法简单快速,故针对大批量样本抽提时可有效降低成本.%Elateridae insects have various species and wide distribution,and many of them are principal underground pests.Due to the high similarity of the external morphology of elateridae insects,there is great difficulty in the work of traditional morphological classification,and most of the phylogenetic studies on elateridae family are conducted based on molecular marker.Currently,many extraction methods of insect genome will destroy the whole polypide,we had experiment and improved the extraction method of micro-insect genomic DNA,and higher-quality genomic DNA was extracted by the protease K digestion method on the precondition of not disrupting the main identification characteristics of elateridae,with a piece of muscle in the hind foot as the extraction object.PCR amplification and sequencing of the extracted genome were carried out,and the results showed that the genomic DNA extracted by this method could be applied to molecular related experiments.In addition,this method is simple and quick,and it can effectively reduce costs in the extraction of large number of samples.