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A Fast, Simple and Low-cost DNA Extraction Protocol from Common Ants and Beetles for Multiple Molecular Applications

Received: 14 April 2024     Accepted: 27 April 2024     Published: 17 May 2024
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Abstract

The rapid development of molecular biology tools in insect systematics, invasion research, evolutionary ecology and biodiversity analysis has led to faster and greater progress in understanding insect behavior and biology. Efficient DNA extraction is the foremost step and serves as the vital foundation. Several rapid DNA extraction methods have been established, which are often time-consuming and labour-intensive. Here, a simple, fast, low-cost DNA extraction protocol for common insect samples was developed basing on 28 specimens of 16 insect species (7 ants, 9 bark and ambrosia beetles). The new protocol was shown to be feasible and highly efficient by comparison with commercial kit in terms of DNA yield, purity and PCR sensitivity. The concentration of DNA through the new rapid method was higher than that through commercial kit, whether in ant or beetle samples. A better quality of DNA extracted via kit was indicated by A260/A280 mostly ranging from 1.80 to 2.00. There was little difference between DNA extracted from adult and nymphal insects. PCR sensitivity of extracted DNA using both protocols was comparable. For nested PCR, amplification after two rounds yielded a bright signal using template DNA through both methods. But for PCR using primers of LCO1490 and HCO2198, the success ratio was lower (85.18%). Through BLAST, these amplicons were matched to related data with high identity. By combining this protocol with variable analysis platforms such as common PCR, loop-mediated isothermal amplification, and high throughput sequencing, it could assist insect diagnostics, biological surveys and invasion researches.

Published in American Journal of Bioscience and Bioengineering (Volume 12, Issue 3)
DOI 10.11648/j.bio.20241203.12
Page(s) 48-55
Creative Commons

This is an Open Access article, distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution and reproduction in any medium or format, provided the original work is properly cited.

Copyright

Copyright © The Author(s), 2024. Published by Science Publishing Group

Keywords

DNA Extraction, Insects, Molecular Analysis, Rapid, Simple, Low-Cost

References
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  • APA Style

    Jiaying, W., Junxia, C., Li, L., Yuanjing, W., Shuyi, Y., et al. (2024). A Fast, Simple and Low-cost DNA Extraction Protocol from Common Ants and Beetles for Multiple Molecular Applications. American Journal of Bioscience and Bioengineering, 12(3), 48-55. https://doi.org/10.11648/j.bio.20241203.12

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    ACS Style

    Jiaying, W.; Junxia, C.; Li, L.; Yuanjing, W.; Shuyi, Y., et al. A Fast, Simple and Low-cost DNA Extraction Protocol from Common Ants and Beetles for Multiple Molecular Applications. Am. J. BioSci. Bioeng. 2024, 12(3), 48-55. doi: 10.11648/j.bio.20241203.12

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    AMA Style

    Jiaying W, Junxia C, Li L, Yuanjing W, Shuyi Y, et al. A Fast, Simple and Low-cost DNA Extraction Protocol from Common Ants and Beetles for Multiple Molecular Applications. Am J BioSci Bioeng. 2024;12(3):48-55. doi: 10.11648/j.bio.20241203.12

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  • @article{10.11648/j.bio.20241203.12,
      author = {Wang Jiaying and Cui Junxia and Liu Li and Wang Yuanjing and Yan Shuyi and Chen Xianfeng},
      title = {A Fast, Simple and Low-cost DNA Extraction Protocol from Common Ants and Beetles for Multiple Molecular Applications
    },
      journal = {American Journal of Bioscience and Bioengineering},
      volume = {12},
      number = {3},
      pages = {48-55},
      doi = {10.11648/j.bio.20241203.12},
      url = {https://doi.org/10.11648/j.bio.20241203.12},
      eprint = {https://article.sciencepublishinggroup.com/pdf/10.11648.j.bio.20241203.12},
      abstract = {The rapid development of molecular biology tools in insect systematics, invasion research, evolutionary ecology and biodiversity analysis has led to faster and greater progress in understanding insect behavior and biology. Efficient DNA extraction is the foremost step and serves as the vital foundation. Several rapid DNA extraction methods have been established, which are often time-consuming and labour-intensive. Here, a simple, fast, low-cost DNA extraction protocol for common insect samples was developed basing on 28 specimens of 16 insect species (7 ants, 9 bark and ambrosia beetles). The new protocol was shown to be feasible and highly efficient by comparison with commercial kit in terms of DNA yield, purity and PCR sensitivity. The concentration of DNA through the new rapid method was higher than that through commercial kit, whether in ant or beetle samples. A better quality of DNA extracted via kit was indicated by A260/A280 mostly ranging from 1.80 to 2.00. There was little difference between DNA extracted from adult and nymphal insects. PCR sensitivity of extracted DNA using both protocols was comparable. For nested PCR, amplification after two rounds yielded a bright signal using template DNA through both methods. But for PCR using primers of LCO1490 and HCO2198, the success ratio was lower (85.18%). Through BLAST, these amplicons were matched to related data with high identity. By combining this protocol with variable analysis platforms such as common PCR, loop-mediated isothermal amplification, and high throughput sequencing, it could assist insect diagnostics, biological surveys and invasion researches.
    },
     year = {2024}
    }
    

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  • TY  - JOUR
    T1  - A Fast, Simple and Low-cost DNA Extraction Protocol from Common Ants and Beetles for Multiple Molecular Applications
    
    AU  - Wang Jiaying
    AU  - Cui Junxia
    AU  - Liu Li
    AU  - Wang Yuanjing
    AU  - Yan Shuyi
    AU  - Chen Xianfeng
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    T2  - American Journal of Bioscience and Bioengineering
    JF  - American Journal of Bioscience and Bioengineering
    JO  - American Journal of Bioscience and Bioengineering
    SP  - 48
    EP  - 55
    PB  - Science Publishing Group
    SN  - 2328-5893
    UR  - https://doi.org/10.11648/j.bio.20241203.12
    AB  - The rapid development of molecular biology tools in insect systematics, invasion research, evolutionary ecology and biodiversity analysis has led to faster and greater progress in understanding insect behavior and biology. Efficient DNA extraction is the foremost step and serves as the vital foundation. Several rapid DNA extraction methods have been established, which are often time-consuming and labour-intensive. Here, a simple, fast, low-cost DNA extraction protocol for common insect samples was developed basing on 28 specimens of 16 insect species (7 ants, 9 bark and ambrosia beetles). The new protocol was shown to be feasible and highly efficient by comparison with commercial kit in terms of DNA yield, purity and PCR sensitivity. The concentration of DNA through the new rapid method was higher than that through commercial kit, whether in ant or beetle samples. A better quality of DNA extracted via kit was indicated by A260/A280 mostly ranging from 1.80 to 2.00. There was little difference between DNA extracted from adult and nymphal insects. PCR sensitivity of extracted DNA using both protocols was comparable. For nested PCR, amplification after two rounds yielded a bright signal using template DNA through both methods. But for PCR using primers of LCO1490 and HCO2198, the success ratio was lower (85.18%). Through BLAST, these amplicons were matched to related data with high identity. By combining this protocol with variable analysis platforms such as common PCR, loop-mediated isothermal amplification, and high throughput sequencing, it could assist insect diagnostics, biological surveys and invasion researches.
    
    VL  - 12
    IS  - 3
    ER  - 

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