Issue
A simplified and optimized protocol for total DNA extraction from insect species: applicable for studying genetic diversity and PCR-based specimen identification via partial amplification of cytochrome oxidase I (COI) gene
Corresponding Author(s) : Kheirollah Yari
Cellular and Molecular Biology,
Vol. 64 No. 12: Issue 12
Abstract
The efficient DNA extraction from insects has been suggested as a critical and main step affecting molecular entomology for taxonomic identification, the establishment of DNA barcoding library and analysis of genetic diversity relationship between insect populations. For successfully apply these molecular techniques, high-quantity and high-quality of the extracted DNA are required. Several protocols for efficient genomic DNA extraction from insects have been developed. In this research, we represent a rapid, reliable and cost-effective method that it is not reliant on poisonous and enzymatic reagents for DNA extraction from insect tissues. Results showed that high quantity and high-quality of the isolated DNA by this method is suitable and can be used directly for PCR, also is enough to do hundreds of molecular reactions. In conclusion, we described a fast, cost-effective, non-toxic and enzyme-free protocol for high yield genomic DNA extraction from green Lacewings (Chrysoperla carnea) tissues in basic equipment laboratories.
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- Dittrich"Schröder G, Wingfield MJ, Klein H, Slippers B. DNA extraction techniques for DNA barcoding of minute gall"inhabiting wasps. Molecular ecology resources. 2012;12(1):109-15.
- Lagisz M, Port G, Wolff K. A cost"effective, simple and high"throughput method for DNA extraction from insects. Insect science. 2010;17(5):465-70.
- Tanaka R, Kikuchi T, Aikawa T, Kanzaki N. Simple and quick methods for nematode DNA preparation. Applied Entomology and Zoology. 2012;47(3):291-4.
- Winterton S, De Freitas S. Molecular phylogeny of the green lacewings (Neuroptera: Chrysopidae). Austral Entomology. 2006;45(3):235-43.
- Mirmoayedi A, Kahrizi D, Pani S, Yari K. Molecular genetic diversity within Myrmeleontidae family. Molecular biology reports. 2013;40(1):639-43.
- Arimoto M, Satoh M, Uesugi R, Osakabe M. DNA preparation method in eggs, immature stages, and diapausing females of Tetranychus spider mites (Acari: Tetranychidae) for diagnostic PCR-RFLP. Applied entomology and zoology. 2012;47(3):295-300.
- Calderón-Cortés N, Quesada M, Cano-Camacho H, Zavala-Páramo G. A simple and rapid method for DNA isolation from xylophagous insects. International journal of molecular sciences. 2010;11(12):5056-64.
- Amani J, Kazemi R, Abbasi AR, Salmanian AH. A simple and rapid leaf genomic DNA extraction method for polymerase chain reaction analysis. Iranian journal of biotechnology. 2011;9(1):69-71.
- Gupta S, Preet S. Protocol optimization for genomic DNA extraction and RAPD-PCR in mosquito larvae (Diptera: Culicidae). Annals of Biological Research. 2012;3(3):1553-61.
- Shams SS, Vahed SZ, Soltanzad F, Kafil V, Barzegari A, Atashpaz S, et al. Highly effective DNA extraction method from fresh, frozen, dried and clotted blood samples. BioImpacts: BI. 2011;1(3):183.
- Minas K, McEwan NR, Newbold CJ, Scott KP. Optimization of a high-throughput CTAB-based protocol for the extraction of qPCR-grade DNA from rumen fluid, plant and bacterial pure cultures. FEMS microbiology letters. 2011;325(2):162-9.
- Yari K, Afzali S, Mozafari H, Mansouri K, Mostafaie A. Molecular cloning, expression and purification of recombinant soluble mouse endostatin as an anti-angiogenic protein in Escherichia coli. Molecular biology reports. 2013;40(2):1027-33.
- Ayazi G, Pirayesh M, Yari K. Analysis of interleukin-1β gene polymorphism and its association with generalized aggressive periodontitis disease. DNA and cell biology. 2013;32(7):409-13.
- Mirmoayedi A, Kahrizi D, Ebadi AA, Yari K, Mohammadi M. Study of individual and sex genetic diversity among each genus and between two genera of Chrysopa and Chrysoperla (Neuroptera, Chrysopidae) based on RAPD-PCR polymorphism. Molecular biology reports. 2012;39(9):8999-9006.
- Skujienć— G, Soroka M. A comparison of different DNA extraction methods for slugs (Mollusca: Pulmonata). Ekologija. 2003;1:12-6.
- Chen H, Rangasamy M, Tan SY, Wang H, Siegfried BD. Evaluation of five methods for total DNA extraction from western corn rootworm beetles. PLoS One. 2010;5(8):e11963.
- Dale C, Justi SA, Galví£o C. Tropical insect collections and DNA extraction, using Rhodnius Stål 1859 (Hemiptera: Heteroptera: Reduviidae: Triatominae). Zootaxa. 2013;3694(4):398-9.
- Wang Q, Wang X. Comparison of methods for DNA extraction from a single chironomid for PCR analysis. Pak J Zool. 2012;44(2):421-6.
- Oliveira L, Wallau G, Loreto E. Isolation of high quality DNA: a protocol combining "rennet” and glass milk. Electronic Journal of Biotechnology. 2009;12(2):1-6.
- Sahu SK, Thangaraj M, Kathiresan K. DNA extraction protocol for plants with high levels of secondary metabolites and polysaccharides without using liquid nitrogen and phenol. ISRN Molecular Biology. 2012;2012.
- Pereira JC, Chaves R, Bastos E, Leití£o A, Guedes-Pinto H. An efficient method for genomic DNA extraction from different molluscs species. International journal of molecular sciences. 2011;12(11):8086-95.
- Watts PC, Thompson DJ, Allen KA, Kemp SJ. How useful is DNA extracted from the legs of archived insects for microsatellite-based population genetic analyses? Journal of Insect Conservation. 2007;11(2):195-8.
- Harvey ML. An alternative for the extraction and storage of DNA from insects in forensic entomology. Journal of Forensic Science. 2005;50(3):1-3.
References
Dittrich"Schröder G, Wingfield MJ, Klein H, Slippers B. DNA extraction techniques for DNA barcoding of minute gall"inhabiting wasps. Molecular ecology resources. 2012;12(1):109-15.
Lagisz M, Port G, Wolff K. A cost"effective, simple and high"throughput method for DNA extraction from insects. Insect science. 2010;17(5):465-70.
Tanaka R, Kikuchi T, Aikawa T, Kanzaki N. Simple and quick methods for nematode DNA preparation. Applied Entomology and Zoology. 2012;47(3):291-4.
Winterton S, De Freitas S. Molecular phylogeny of the green lacewings (Neuroptera: Chrysopidae). Austral Entomology. 2006;45(3):235-43.
Mirmoayedi A, Kahrizi D, Pani S, Yari K. Molecular genetic diversity within Myrmeleontidae family. Molecular biology reports. 2013;40(1):639-43.
Arimoto M, Satoh M, Uesugi R, Osakabe M. DNA preparation method in eggs, immature stages, and diapausing females of Tetranychus spider mites (Acari: Tetranychidae) for diagnostic PCR-RFLP. Applied entomology and zoology. 2012;47(3):295-300.
Calderón-Cortés N, Quesada M, Cano-Camacho H, Zavala-Páramo G. A simple and rapid method for DNA isolation from xylophagous insects. International journal of molecular sciences. 2010;11(12):5056-64.
Amani J, Kazemi R, Abbasi AR, Salmanian AH. A simple and rapid leaf genomic DNA extraction method for polymerase chain reaction analysis. Iranian journal of biotechnology. 2011;9(1):69-71.
Gupta S, Preet S. Protocol optimization for genomic DNA extraction and RAPD-PCR in mosquito larvae (Diptera: Culicidae). Annals of Biological Research. 2012;3(3):1553-61.
Shams SS, Vahed SZ, Soltanzad F, Kafil V, Barzegari A, Atashpaz S, et al. Highly effective DNA extraction method from fresh, frozen, dried and clotted blood samples. BioImpacts: BI. 2011;1(3):183.
Minas K, McEwan NR, Newbold CJ, Scott KP. Optimization of a high-throughput CTAB-based protocol for the extraction of qPCR-grade DNA from rumen fluid, plant and bacterial pure cultures. FEMS microbiology letters. 2011;325(2):162-9.
Yari K, Afzali S, Mozafari H, Mansouri K, Mostafaie A. Molecular cloning, expression and purification of recombinant soluble mouse endostatin as an anti-angiogenic protein in Escherichia coli. Molecular biology reports. 2013;40(2):1027-33.
Ayazi G, Pirayesh M, Yari K. Analysis of interleukin-1β gene polymorphism and its association with generalized aggressive periodontitis disease. DNA and cell biology. 2013;32(7):409-13.
Mirmoayedi A, Kahrizi D, Ebadi AA, Yari K, Mohammadi M. Study of individual and sex genetic diversity among each genus and between two genera of Chrysopa and Chrysoperla (Neuroptera, Chrysopidae) based on RAPD-PCR polymorphism. Molecular biology reports. 2012;39(9):8999-9006.
Skujienć— G, Soroka M. A comparison of different DNA extraction methods for slugs (Mollusca: Pulmonata). Ekologija. 2003;1:12-6.
Chen H, Rangasamy M, Tan SY, Wang H, Siegfried BD. Evaluation of five methods for total DNA extraction from western corn rootworm beetles. PLoS One. 2010;5(8):e11963.
Dale C, Justi SA, Galví£o C. Tropical insect collections and DNA extraction, using Rhodnius Stål 1859 (Hemiptera: Heteroptera: Reduviidae: Triatominae). Zootaxa. 2013;3694(4):398-9.
Wang Q, Wang X. Comparison of methods for DNA extraction from a single chironomid for PCR analysis. Pak J Zool. 2012;44(2):421-6.
Oliveira L, Wallau G, Loreto E. Isolation of high quality DNA: a protocol combining "rennet” and glass milk. Electronic Journal of Biotechnology. 2009;12(2):1-6.
Sahu SK, Thangaraj M, Kathiresan K. DNA extraction protocol for plants with high levels of secondary metabolites and polysaccharides without using liquid nitrogen and phenol. ISRN Molecular Biology. 2012;2012.
Pereira JC, Chaves R, Bastos E, Leití£o A, Guedes-Pinto H. An efficient method for genomic DNA extraction from different molluscs species. International journal of molecular sciences. 2011;12(11):8086-95.
Watts PC, Thompson DJ, Allen KA, Kemp SJ. How useful is DNA extracted from the legs of archived insects for microsatellite-based population genetic analyses? Journal of Insect Conservation. 2007;11(2):195-8.
Harvey ML. An alternative for the extraction and storage of DNA from insects in forensic entomology. Journal of Forensic Science. 2005;50(3):1-3.