TY - JOUR
T1 - Comparison of 3 DNA extraction methods for extracting DNA from an adult Culex quinquefasciatus (Diptera: Culicidae)
AU - Firmansyah, Nurhadi Eko
AU - Thongseesuksai, Thaksaporn
AU - Boonmars, Thidarut
AU - Sungkar, Saleha
AU - Laummaunwai, Porntip
N1 - Publisher Copyright:
© The Author(s) 2023. Published by Oxford University Press on behalf of Entomological Society of America.
PY - 2023/9/1
Y1 - 2023/9/1
N2 - 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.
AB - 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.
KW - cost- and time-effective
KW - DNAzol Direct reagent
KW - microwave irradiation
KW - mosquito
KW - polymerase chain reaction
UR - http://www.scopus.com/inward/record.url?scp=85174917723&partnerID=8YFLogxK
U2 - 10.1093/jisesa/iead080
DO - 10.1093/jisesa/iead080
M3 - Article
C2 - 37804500
AN - SCOPUS:85174917723
SN - 1536-2442
VL - 23
JO - Journal of Insect Science
JF - Journal of Insect Science
IS - 5
M1 - iead080
ER -