| 一种蚜茧蜂基因组DNA提取方法 |
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| 引用本文： | 孔博,张宏瑞,和淑琪,谷星慧,杨硕媛,张立猛,李正跃.一种蚜茧蜂基因组DNA提取方法[J].中国烟草学报,2017,23(6):88-93. |
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| 作者姓名： | 孔博 张宏瑞 和淑琪 谷星慧 杨硕媛 张立猛 李正跃 |
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| 作者单位： | 1 云南农业大学植物保护学院, 云南昆明 650201; |
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| 基金项目： | 云南农业大学研究生科技创新项目（2016ykc16），中国烟草总公司科技重大专项项目（110201601021） |
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| 摘 要： | 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。
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| 关 键 词： | 蚜茧蜂 基因组DNA DNA提取 PCR |
| 收稿时间： | 2017-03-23 |
Method for extraction of genomic DNA from Aphidiidae |
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| Affiliation: | 1 Plant Protection College, Yunnan Agricultural University, Kunming 650201, China;2 Yunnan Yuxi Municipal Tobacco Company, Yuxi 653100, Yunnan, China |
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| Abstract: | In order to effectively extract genomic DNA of Aphidiidae and reserve specimens for morphological identification, 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 verified by amplification of mitochondrial 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 identification. 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 amplified bands were clear and complete. |
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