Department of Parasitology, Faculty of Medicine, Khon Kaen University, Khon Kaen, Thailand.
Neglected, Zoonosis and Vector-Borne Disease Research Group, Khon Kaen University, Khon Kaen, Thailand.
J Insect Sci. 2023 Sep 1;23(5). doi: 10.1093/jisesa/iead080.
The aim of this study was to compare 3 DNA extraction methods: the PureLink Genomic DNA kit, DNAzol Direct reagent, and a microwave-based method, for extracting DNA from an adult Culex quinquefasciatus by focusing on the quantity and purity of DNA, cost, and time required. Ten mosquitoes were individually used for DNA extraction by each method. Based on the results obtained, DNA was extracted from each method using specific primers, resulting in a polymerase chain reaction (PCR) product with a length of 274 bp. The DNA quantity extracted using the DNAzol Direct (179.08 ± 3.77 ng/µl) differs significantly from that of the commercial kit (115.98 ± 4.57 ng/µl) and a microwave-based method (119.26 ± 3.06 ng/µl). The absorbance ratio of DNA extracted using the PureLink Genomic DNA kit, the DNAzol Direct, and the microwave-based methods was 1.92 ± 0.02, 1.79 ± 0.01, and 1.87 ± 0.01, respectively. Among the 3 methods evaluated, the microwave-based method is simpler, less expensive, and more time efficient. This is the first evaluation of the microwave-based method for extracting DNA from an adult mosquito. This study provides a useful guide for alternative DNA extraction methods for PCR-based assays, especially in low-resource settings.
本研究旨在比较 3 种 DNA 提取方法:PureLink Genomic DNA 试剂盒、DNAzol Direct 试剂和微波法,重点关注 DNA 的提取量和纯度、成本和所需时间,从成年致倦库蚊中提取 DNA。每种方法各用 10 只蚊子进行 DNA 提取。根据结果,使用特定引物从每种方法中提取 DNA,得到长度为 274bp 的聚合酶链反应(PCR)产物。DNAzol Direct 提取的 DNA 量(179.08 ± 3.77ng/µl)与商用试剂盒(115.98 ± 4.57ng/µl)和微波法(119.26 ± 3.06ng/µl)差异显著。使用 PureLink Genomic DNA 试剂盒、DNAzol Direct 和微波法提取的 DNA 的吸光度比值分别为 1.92 ± 0.02、1.79 ± 0.01 和 1.87 ± 0.01。在评价的 3 种方法中,微波法更简单、更便宜、更省时。这是首次评估微波法从成年蚊子中提取 DNA。本研究为基于 PCR 的检测提供了一种有用的替代 DNA 提取方法指南,特别是在资源有限的环境中。