| 摘要: |
| 海绵标本长期、完整的保存是开展海绵分类学及分子进化研究的重要前提, 本研究以建立通用的海绵动物标本的保存方法及快速、完整的基因组DNA 提取方法为主要目的, 通过对硅质的山海绵(Mycale sp.)和钙质的白枝海绵(Leucosoleniidae sp.)为材料进行–20℃冰冻、乙醇固定、冰冻后风干和乙醇固定后风干等4 种保存方法进行研究, 同时采用酚-氯仿法、高盐法、CTAB 法等3 种基因组DNA提取方法, 测试了4 种保存方法的DNA 提取效率。实验结果显示, 海绵样品的4 种保存方法以及3 种DNA 提取方法对于硅质海绵以及钙质海绵虽然在提取的DNA 得率上有差异, 但都能获得较高纯度基因组DNA。从经济成本、方便性、潜在的污染等因素考虑, 高盐法是首选的提取海绵基因组DNA 的方法; 乙醇固定保存的海绵样品DNA 得率最高, 冰冻的海绵样品得率最低, 推荐采用乙醇固定保存海绵样品的方法。 |
| 关键词: 海绵 DNA 提取 保存 |
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| 基金项目:中央高校基本科研业务费专项资金资助项目(2010121034);福建省自然科学基金资助项目(2011J01245) |
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| Comparison of samples preservation and DNA extraction of sponges |
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| Abstract: |
| In order to build the optional method of sponge preservation and a safe, fast and effective genomic DNA extraction method, different ways of DNA extraction and preservation were studied in this paper. Two different species, Mycale sp. and Leucosoleniidae sp., and four different ways of preservation including ?20℃ freezing, ethanol fixation, air drying and ethanol fixation then drying were tested. Three methods for DNA extraction including cetyltrimethylammonium bromide (CTAB), Phenol – Chloroform and high salt extraction were examined. The results show that each method can obtain high quality genomic DNA. But considering economical, enviromental friendly and convenient factors, the method of high salt extraction was the best one among the three methods of DNA extraction. The amount and purity of DNA isolated from ethanol fixed specimens were the best. In a word, the ethanol fixed is the best method of sponge preservation. |
| Key words: Sponge genomic DNA extraction preservation |
