一种蚜茧蜂基因组DNA提取方法

摘要

In order to eff ectively extract genomic DNA of Aphidiidae and reserve specimens for morphological identifi cation, extraction method of conventional kits (column centrifugation method) was improved. Genomic DNA of 7 species of Aphidiidae was extracted by puncturing,and genomic DNA of 4 species among them were extracted by grinding. Purity and yield of genomic DNA extracted by two methods were compared and verifi ed by amplifi cation of m itochondrial COI gene sequences. Results showed that puncturing extraction could be used to extract genomic DNA from individual adult of Ahidiidae, and to reserve specimens for morphological identifi cation. The yield of DNA extracted by this method ranged from 29 to 55 ng/μL,OD260 /OD280 of the extracts ranged from 1.51 to 1.91, which could stably amplify mitochondrial COI gene sequences, and PCR amplifi ed bands were clear and complete.%为有效获得蚜茧蜂基因组DNA,并保留其虫体做形态鉴定.本文对常规试剂盒提取方法(柱离心法)进行改进,采用穿刺法提取7种蚜茧蜂的基因组DNA,同时以研磨法提取4种常见蚜茧蜂成虫基因组DNA做对照,比较两种方法提取的基因组DNA的纯度和产量,并通过线粒体COI基因序列扩增进行验证.结果表明:穿刺法能从单头蚜茧蜂中提取DNA而不影响形态鉴定.该方法提取的DNA产物产量在29~55 ng/μL,提取物的OD260/OD280在1.51~1.91,可以稳定地进行线粒体COI基因序列扩增,PCR扩增条带清晰完整.