基于磁珠的蚊虫 DNA 提取方案用于下一代测序。

A Magnetic-Bead-Based Mosquito DNA Extraction Protocol for Next-Generation Sequencing.

机构信息

Florida Medical Entomology Laboratory, Department of Entomology and Nematology, Institute of Food and Agricultural Sciences, University of Florida.

U. S. Fish and Wildlife Service, Pacific Islands Fish and Wildlife Office.

出版信息

J Vis Exp. 2021 Apr 15(170). doi: 10.3791/62354.

DOI:10.3791/62354

PMID:33938890

Abstract

A recently published DNA extraction protocol using magnetic beads and an automated DNA extraction instrument suggested that it is possible to extract high quality and quantity DNA from a well-preserved individual mosquito sufficient for downstream whole genome sequencing. However, reliance on an expensive automated DNA extraction instrument can be prohibitive for many laboratories. Here, the study provides a budget-friendly magnetic-bead-based DNA extraction protocol, which is suitable for low to medium throughput. The protocol described here was successfully tested using individual Aedes aegypti mosquito samples. The reduced costs associated with high quality DNA extraction will increase the application of high throughput sequencing to resource limited labs and studies.

摘要

最近发表的一项 DNA 提取方案，使用了磁珠和自动化 DNA 提取仪器，表明从保存完好的个体蚊子中提取高质量和数量的 DNA 是可行的，足以用于下游的全基因组测序。然而，对于许多实验室来说，依赖昂贵的自动化 DNA 提取仪器可能是不可行的。在这里，本研究提供了一种经济实惠的基于磁珠的 DNA 提取方案，适用于低到中等通量。这里描述的方案已成功应用于单个埃及伊蚊样本。高质量 DNA 提取的成本降低将增加高通量测序在资源有限的实验室和研究中的应用。