Extracting DNA from Preserved Mosquito Blood Meals.

Lawrence E Reeves, Nathan D Burkett-Cadena
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引用次数: 0

Abstract

Mosquito species vary in their host associations. Although some species are relative generalists, most specialize, to varying extents, on particular types of host animals. Mosquito host associations are among the most important factors that influence the transmission dynamics of mosquito-vectored pathogens, and understanding these associations can provide insight on how such pathogens move within ecosystems. Characterization of the host associations of mosquito species requires applying blood meal analysis to the largest possible sample size of mosquito blood meals. Processing large samples of mosquito blood meals can be time-consuming, especially when chain-termination sequencing is used, necessitating individual processing of each specimen. Various methods and commercially available kits and products are available for extracting DNA from mosquito blood meals. The hot sodium hydroxide and Tris (HotSHOT) method is a rapid and inexpensive method of DNA extraction that is compatible with the recovery of DNA from mosquito blood meals preserved on QIAcard Flinders Technology Associates (FTA) Classic Cards (FTA cards). FTA cards allow nucleic acids found in blood meals to be preserved easily, even in field conditions. DNA prepared using this method is suitable for polymerase chain reaction (PCR)-based blood meal analysis.

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从保存的蚊子血餐中提取 DNA。
蚊子物种与宿主的关系各不相同。虽然有些物种是相对的通食动物,但大多数物种都在不同程度上专攻特定类型的宿主动物。蚊子的宿主关联是影响蚊子传播的病原体传播动态的最重要因素之一,了解这些关联可以深入了解这些病原体如何在生态系统中移动。要确定蚊子物种宿主关联的特征,需要对尽可能多的蚊子血餐样本进行血餐分析。处理大量蚊子血餐样本非常耗时,尤其是使用链终止测序法时,必须对每个样本进行单独处理。从蚊子血餐中提取 DNA 有多种方法和市售试剂盒及产品。热氢氧化钠和三羟色胺(HotSHOT)法是一种快速、廉价的 DNA 提取方法,可用于从保存在 QIAcard Flinders Technology Associates(FTA)经典卡(FTA 卡)上的蚊子血餐中提取 DNA。即使在野外条件下,FTA 卡也能轻松保存血餐中的核酸。用这种方法制备的 DNA 适用于基于聚合酶链反应 (PCR) 的血餐分析。
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来源期刊
Cold Spring Harbor protocols
Cold Spring Harbor protocols Biochemistry, Genetics and Molecular Biology-Biochemistry, Genetics and Molecular Biology (all)
CiteScore
3.00
自引率
0.00%
发文量
163
期刊介绍: Cold Spring Harbor Laboratory is renowned for its teaching of biomedical research techniques. For decades, participants in its celebrated, hands-on courses and users of its laboratory manuals have gained access to the most authoritative and reliable methods in molecular and cellular biology. Now that access has moved online. Cold Spring Harbor Protocols is an interdisciplinary journal providing a definitive source of research methods in cell, developmental and molecular biology, genetics, bioinformatics, protein science, computational biology, immunology, neuroscience and imaging. Each monthly issue details multiple essential methods—a mix of cutting-edge and well-established techniques.
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