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

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 indivi...

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Autores principales: Firmansyah, Nurhadi Eko, Thongseesuksai, Thaksaporn, Boonmars, Thidarut, Sungkar, Saleha, Laummaunwai, Porntip
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Oxford University Press 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10560001/
https://www.ncbi.nlm.nih.gov/pubmed/37804500
http://dx.doi.org/10.1093/jisesa/iead080
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author Firmansyah, Nurhadi Eko
Thongseesuksai, Thaksaporn
Boonmars, Thidarut
Sungkar, Saleha
Laummaunwai, Porntip
author_facet Firmansyah, Nurhadi Eko
Thongseesuksai, Thaksaporn
Boonmars, Thidarut
Sungkar, Saleha
Laummaunwai, Porntip
author_sort Firmansyah, Nurhadi Eko
collection PubMed
description 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.
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spelling pubmed-105600012023-10-08 Comparison of 3 DNA extraction methods for extracting DNA from an adult Culex quinquefasciatus (Diptera: Culicidae) Firmansyah, Nurhadi Eko Thongseesuksai, Thaksaporn Boonmars, Thidarut Sungkar, Saleha Laummaunwai, Porntip J Insect Sci Research 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. Oxford University Press 2023-10-07 /pmc/articles/PMC10560001/ /pubmed/37804500 http://dx.doi.org/10.1093/jisesa/iead080 Text en © The Author(s) 2023. Published by Oxford University Press on behalf of Entomological Society of America. https://creativecommons.org/licenses/by-nc-nd/4.0/This is an Open Access article distributed under the terms of the Creative Commons Attribution-NonCommercial-NoDerivs licence (https://creativecommons.org/licenses/by-nc-nd/4.0/), which permits non-commercial reproduction and distribution of the work, in any medium, provided the original work is not altered or transformed in any way, and that the work is properly cited. For commercial re-use, please contact journals.permissions@oup.com
spellingShingle Research
Firmansyah, Nurhadi Eko
Thongseesuksai, Thaksaporn
Boonmars, Thidarut
Sungkar, Saleha
Laummaunwai, Porntip
Comparison of 3 DNA extraction methods for extracting DNA from an adult Culex quinquefasciatus (Diptera: Culicidae)
title Comparison of 3 DNA extraction methods for extracting DNA from an adult Culex quinquefasciatus (Diptera: Culicidae)
title_full Comparison of 3 DNA extraction methods for extracting DNA from an adult Culex quinquefasciatus (Diptera: Culicidae)
title_fullStr Comparison of 3 DNA extraction methods for extracting DNA from an adult Culex quinquefasciatus (Diptera: Culicidae)
title_full_unstemmed Comparison of 3 DNA extraction methods for extracting DNA from an adult Culex quinquefasciatus (Diptera: Culicidae)
title_short Comparison of 3 DNA extraction methods for extracting DNA from an adult Culex quinquefasciatus (Diptera: Culicidae)
title_sort comparison of 3 dna extraction methods for extracting dna from an adult culex quinquefasciatus (diptera: culicidae)
topic Research
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10560001/
https://www.ncbi.nlm.nih.gov/pubmed/37804500
http://dx.doi.org/10.1093/jisesa/iead080
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