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

Nurhadi Eko Firmansyah, Thaksaporn Thongseesuksai, Thidarut Boonmars, Saleha Sungkar, Porntip Laummaunwai

Research output: Contribution to journalArticlepeer-review

Abstract

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.

Original languageEnglish
Article numberiead080
JournalJournal of Insect Science
Volume23
Issue number5
DOIs
Publication statusPublished - 1 Sept 2023

Keywords

  • cost- and time-effective
  • DNAzol Direct reagent
  • microwave irradiation
  • mosquito
  • polymerase chain reaction

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