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From the Field to the Lab: Best Practices for Field Preservation of Bat Specimens for Molecular Analyses

机译:从野外到实验室:蝙蝠标本在野外保存的最佳实践用于分子分析

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摘要

Studies in molecular ecology depend on field-collected samples for genetic information, and the tissue sampled and preservation conditions strongly affect the quality of the DNA obtained. DNA yields from different tissue types have seldom been compared, and the relative performance of storage media has never been directly tested, even though these media may influence DNA degradation under field conditions. We analyzed DNA yield from buccal swabs and wing punches harvested from live bats using nucleic acid quantification as well as quantitative PCR for a single-copy nuclear locus. We also compared DNA yields from wing tissue preserved in three media: ethanol, NaCl-saturated dimethyl sulfoxide (DMSO), and silica desiccant. Wing punches yielded more total DNA than did buccal swabs, and wing tissues preserved in silica beads yielded significantly more total and nuclear DNA than those preserved in DMSO or ethanol. These results show that tissue type and preservation media strongly influence the quantity of DNA obtained from non-lethal genetic samples, and based on these effects we provide recommendations for field collection of tissues for genetic analyses.
机译:分子生态学的研究依赖于现场采集的样本来获取遗传信息,而采样的组织和保存条件会极大地影响获得的DNA的质量。很少比较来自不同组织类型的DNA产量,并且从未直接测试存储介质的相对性能,即使这些介质可能会影响田间条件下的DNA降解。我们使用核酸定量以及定量PCR的单拷贝核基因座分析了从活蝙蝠采集的颊拭子和翼拳的DNA产量。我们还比较了保存在三种介质中的机翼组织的DNA产量:乙醇,NaCl饱和的二甲基亚砜(DMSO)和二氧化硅干燥剂。拳打产生的总DNA比颊拭子产生的DNA多,而保存在硅胶珠中的翅膀组织产生的总DNA和核DNA明显多于DMSO或乙醇。这些结果表明,组织类型和保存介质会强烈影响从非致死性遗传样品中获得的DNA数量,并且基于这些影响,我们为在现场收集组织进行遗传分析提供了建议。

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