| 摘要: |
| 对有孔虫的DNA进行有效保存及提取是开展有孔虫分子鉴定工作的前提。本研究以探索适合玻璃质壳类底栖有孔虫的DNA保存及提取方法为目的, 以中国近海优势属——玻璃质类底栖有孔虫卷转虫属(Ammonia spp.)为模式生物, 考察不同温度(20-120℃, 间隔10℃)烘干、冷冻(-20℃、-80℃, 有水、无水)以及脱氧胆酸钠裂解液(Na deoxycholate, 简称DOC)处理(含和不含EDTA)保存有孔虫DNA方法, 同时进行了DOC裂解液法和Guanidine裂解液法对Ammonia spp. DNA提取效能的比较。结果显示, 烘干保存法中20℃组和30℃组显著优于40℃组(P<0.05); 冷冻保存法中无水组显著优于有水组(P<0.05); DOC裂解液保存法各组间无显著差异(P>0.05)。三种保存方法综合效果表现为冷冻组显著优于烘干组和裂解液组(P<0.05); 两种DNA提取方法的提取效果都很好, 但DOC法更廉价且操作简便。本文给出了一套适用于玻璃质壳类有孔虫的DNA保存和提取的优化方案。 |
| 关键词: 玻璃质壳 底栖有孔虫 分子生物学 Ammonia DNA提取 保存 |
| DOI：10.11693/hyhz20150400097 |
| 分类号: |
| 基金项目:国家自然科学基金项目, 41476043号, 41230959号, 41176132号; 中国科学院战略性先导科技专项(A类)项目, XDA11030104号; 大陆架科学钻探项目, GZH201100202号; 国家海洋局全球变化与海气相互作用专项项目, GASI-03-01-03-01号, DOMEP(MEA)-01-01-E号; 中华人民共和国科学技术部项目, 2014FY110500号。 |
附件 |
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| EFFICIENCY OF DNA PRESERVATION AND EXTRACTION FROM BENTHIC HYALINE FORAMINIFERA OF AMMONIA SPP.: A METHODOLOGICAL COMPARISON |
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LYU Man1,2, LEI Yan-Li1, LI Tie-Gang1,3
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1.Department of Marine Organism Taxonomy & Phylogeny, Institute of Oceanology, Chinese Academy of Sciences, Qingdao 266071, China;2.University of Chinese Academy of Sciences, Beijing 100049, China;3.Key Laboratory of Marine Sedimentology and Environmental Geology, First Institute of Oceanography, SOA, Laboratory for Marine Geology, Qingdao National Laboratory for Marine Science and Technology, Qingdao 266061, China
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| Abstract: |
| In order to establish efficient methods for extracting and preserving DNA from hyaline benthic foraminifera Ammonia spp., two experimental series of DNA preservation were set up including (1) air-dried (at 20, 30, 40, 50, 60, 70, 80, 90, 100, 110, and 120℃) vs frozen (-20 and -80℃, with or without seawater); (2) DOC lysis buffer with vs. without EDTA. In addition, two extraction methods for DNA (DOC lysis buffer and Guanidine lysis buffer) were compared. The results show that the DNA was better preserved under the condition of air-dried under 20℃ and 30℃ than 40℃ (P<0.05); frozen without seawater group was better than with seawater group (P<0.05). There was no significant difference between the DOC lysis buffer with or without EDTA treatment (P>0.05). However, the optimized condition was obtained in the treatment with frozen samples. Therefore, the two DNA extraction methods are both effective, but DOC lysis method is more economic and convenient. In the last, optimal DNA-preservation and -extraction protocols are summarized. |
| Key words: hyaline benthic foraminifera molecular biology Ammonia genomic DNA extraction preservation |
