Comparison of 3 DNA extraction methods for extracting DNA from an adult Culex quinquefasciatus (Diptera: Culicidae)

被引:3
作者
Firmansyah, Nurhadi Eko [1 ]
Thongseesuksai, Thaksaporn [1 ]
Boonmars, Thidarut [1 ,2 ]
Sungkar, Saleha [3 ]
Laummaunwai, Porntip [1 ,2 ]
机构
[1] Khon Kaen Univ, Fac Med, Dept Parasitol, Khon Kaen, Thailand
[2] Khon Kaen Univ, Zoonosis & Vector Borne Dis Res Grp, Khon Kaen, Thailand
[3] Univ Indonesia, Fac Med, Dept Parasitol, Jakarta, Indonesia
关键词
mosquito; cost- and time-effective; polymerase chain reaction; microwave irradiation; DNAzol Direct reagent; DIRECT PCR; BACTERIA; COMPLEX;
D O I
10.1093/jisesa/iead080
中图分类号
Q96 [昆虫学];
学科分类号
摘要
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/mu l) differs significantly from that of the commercial kit (115.98 +/- 4.57 ng/mu l) and a microwave-based method (119.26 +/- 3.06 ng/mu l). 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. Graphical Abstract
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页数:7
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