一种简单高效的浮游动物总基因组 DNA 提取方法。

Q1 Multidisciplinary SpringerPlus Pub Date : 2016-12-01 eCollection Date: 2016-01-01 DOI:10.1186/s40064-016-3724-x
Hanafiah Fazhan, Khor Waiho, Md Sheriff Shahreza
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引用次数: 0

摘要

分子方法被广泛应用于微小浮游动物的物种鉴定和分类研究。桡足亚纲是目前最受关注的浮游动物之一。通过分子分析准确鉴定小型桡足类各生命阶段的物种非常重要,尤其是在对桡足类种群进行分类和系统评估以及了解其在海洋群落中的动态方面。然而,由于桡足类桡足动物个体较小,且具有底栖行为,因此从它们身上提取总基因组 DNA(TGDNA)可能存在问题。在这项研究中,对六种针对单个桡足类的 TGDNA 提取方法进行了比较。研究人员设计了第一种简单、可行、高效和一致的 TGDNA 提取新方法,并将其与商业试剂盒和经改良的现有 TGDNA 提取方法进行了比较。与其他方法相比,新描述的 TGDNA 提取方法 "在 PCR 缓冲液中孵育 "法的 PCR 扩增成功率高(82%),结果良好且一致。由于 "PCR 缓冲液中的孵育 "法相对稳定且经济,因此强烈推荐用于其他微小浮游动物的 TGDNA 提取。
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A simple and efficient total genomic DNA extraction method for individual zooplankton.

Molecular approaches are widely applied in species identification and taxonomic studies of minute zooplankton. One of the most focused zooplankton nowadays is from Subclass Copepoda. Accurate species identification of all life stages of the generally small sized copepods through molecular analysis is important, especially in taxonomic and systematic assessment of harpacticoid copepod populations and to understand their dynamics within the marine community. However, total genomic DNA (TGDNA) extraction from individual harpacticoid copepods can be problematic due to their small size and epibenthic behavior. In this research, six TGDNA extraction methods done on individual harpacticoid copepods were compared. The first new simple, feasible, efficient and consistent TGDNA extraction method was designed and compared with the commercial kit and modified available TGDNA extraction methods. The newly described TGDNA extraction method, "Incubation in PCR buffer" method, yielded good and consistent results based on the high success rate of PCR amplification (82%) compared to other methods. Coupled with its relatively consistent and economical method the "Incubation in PCR buffer" method is highly recommended in the TGDNA extraction of other minute zooplankton species.

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来源期刊
SpringerPlus
SpringerPlus MULTIDISCIPLINARY SCIENCES-
CiteScore
1.76
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0.00%
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0
期刊介绍: Cessation
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