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Comparison of DNA extraction protocols for PCR-based techniques in wheat

Yıl 2019, Sayı: 17, 860 - 865, 31.12.2019
https://doi.org/10.31590/ejosat.646491

Öz

Molecular biology
and genetic studies requires high quality DNA isolated from plants. Therefore,
isolation of pure and high quality DNA in a short time is important. However,
since different plants have different chemical composition, in even close species
need specific genomic DNA isolation protocol. There are a large number of
protocols for wheat. Therefore, which method is more applicable, safer and easier
should be determined. For this purpose three DNA extraction protocols were used.
The DNA samples extracted from these protocols were compared for their purity and
concentrations. The quality and amount of DNA obtained was determined by NanoDrop
Spectrophotometer (A260/280 absorbance) and agarose gel electrophoresis. Additionally
DNAs extracted were amplified by polymerase chain reactions (PCR) and the suitability
of isolated DNAs for PCR reactions were also determined. The results show that with
the CTAB DNA extraction methods were extracted the highest DNA concentrations
in wheat (Triticum avestivum L.) compared to other protocols.

Kaynakça

  • Abdel-Latif, A., & Osman, G. (2017). Comparison of three genomic DNA extraction methods to obtain high DNA quality from maize. Plant Methods, 13(1), 1.
  • Abouseadaa, H. H., Osman, G. H., Ramadan, A. M., Hassanein, S. E., Abdelsattar, M. T., Morsy, Y. B., & Gad, A. A. (2015). Development of transgenic wheat (Triticum aestivum L.) expressing avidin gene conferring resistance to stored product insects. BMC plant biology, 15(1), 183.
  • Aljanabi, S. M., & Martinez, I. (1997). Universal and rapid salt-extraction of high quality genomic DNA for PCR-based techniques. Nucleic acids research, 25(22), 4692-4693.Chai, J. F., Liu, X., & Jia, J. Z. (2006). A rapid isolation method of wheat DNA suitable for PCR analysis. Journal of Plant Genetic Resources, 7(2), 246-248.
  • Cristina, D., Ciuca, M., & Cornea, C. P. (2017). Comparison of four genomic DNA isolation methods from single dry seed of wheat, barley and rye. AgroLife Scientific Journal, 6(1), 84-91.
  • Dilworth, E., & Frey, J. E. (2000). A rapid method for high throughput DNA extraction from plant material for PCR amplification. Plant Molecular Biology Reporter, 18(1), 61-64.
  • Doyle J.J., & Doyle J. L. (1987). A rapid DNA isolation procedure for small quantities of fresh leaf tissue, Phytochem. Bull., 9, 11-15.
  • Kadry, A. K., Gamal, O., & Waeil, A. A. (2012). Genetic diversity and taxonomic relationships of some'Ipomoea'species based on analysis of RAPD-PCR and SDS-PAGE of seed proteins. Australian Journal of Crop Science, 6(6), 1088.
  • Khanuja, S. P., Shasany, A. K., Darokar, M. P., & Kumar, S. (1999). Rapid isolation of DNA from dry and fresh samples of plants producing large amounts of secondary metabolites and essential oils. Plant Molecular Biology Reporter, 17(1), 74-74.
  • Lee, P. Y., Costumbrado, J., Hsu, C. Y., Kim, Y. H. (2012). Agarose gel electrophoresis for the separation of DNA fragments. Journal of Visualized Experiments, (62), 3923. Moreira, P. A., & Oliveira, D. A. (2011). Leaf age affects the quality of DNA extracted from Dimorphandra mollis (Fabaceae), a tropical tree species from the Cerrado region of Brazil. Genetics and Molecular Research, 10(1), 353-358.
  • Murray, M. G., & Thompson, W. F. (1980). Rapid isolation of high molecular weight plant DNA. Nucleic acids research, 8(19), 4321-4326.
  • Paris, M., & Carter, M. (2000). Cereal DNA: a rapid high-throughput extraction method for marker assisted selection. Plant Molecular Biology Reporter, 18(4), 357-360.
  • Plaschke, J., Ganal, M. W., & Röder, M. S. (1995). Detection of genetic diversity in closely related bread wheat using microsatellite markers. Theoretical and Applied Genetics, 91(6-7), 1001-1007.
  • Pervaiz, Z. H., Turi, N. A., Khaliq, I., Rabbani, M. A., & Malik, S. A. (2011). Methodology: a modified method for high-quality DNA extraction for molecular analysis in cereal plants. Genetics and molecular research: GMR, 10(3), 1669-1673.
  • Röder, M. S., Plaschke, J., König, S. U., Börner, A., Sorrells, M. E., Tanksley, S. D., & Ganal, M. W. (1995). Abundance, variability and chromosomal location of microsatellites in wheat. Molecular and General Genetics MGG, 246(3), 327-333.
  • Sahu, S. K., Thangaraj, M., & Kathiresan, K. (2012). DNA extraction protocol for plants with high levels of secondary metabolites and polysaccharides without using liquid nitrogen and phenol. ISRN Molecular Biology, 2012.
  • Somers, D. J., Isaac, P., & Edwards, K. (2004). A high-density microsatellite consensus map for bread wheat (Triticum aestivum L.). Theoretical and Applied Genetics, 109(6), 1105-1114.
  • Sönmezoğlu, Ö. A., & Keskin, H. (2015). Determination of genetically modified corn and soy in processed food products. Journal of Applied Biology & Biotechnology, 3, 32-37.
  • Sönmezoğlu, Ö. A., & Terzi, B. (2018). Characterization of some bread wheat genotypes using molecular markers for drought tolerance. Physiology and molecular biology of plants, 24(1), 159-166.
  • Varma, A., Padh, H., & Shrivastava, N. (2007). Plant genomic DNA isolation: an art or a science. Biotechnology Journal: Healthcare Nutrition Technology, 2(3), 386-392.
  • Yadav, M. K., & Chand, P. (2018). Assessment of Genetic Diversity among Twenty Indian Wheat (Triticum aestivum L.) Cultivars using Simple Sequence Repeat (SSR) Markers. Int. J. Curr. Microbiol. App. Sci, 7(3), 1708-1717.

Buğdayda PCR'a dayalı teknikler için DNA ekstraksiyon protokollerinin karşılaştırılması

Yıl 2019, Sayı: 17, 860 - 865, 31.12.2019
https://doi.org/10.31590/ejosat.646491

Öz

Moleküler biyoloji ve genetik çalışmalar, bitkilerden izole edilmiş yüksek kaliteli DNA gerektirir. Bu nedenle, saf ve yüksek kalitede DNA'nın kısa sürede izolasyonu önemlidir. Bununla birlikte, farklı bitkiler farklı kimyasal bileşime sahip olduklarından, yakın türlerde bile spesifik genomik DNA izolasyon protokolüne ihtiyaç vardır. Buğday için çok sayıda protokol vardır. Bu nedenle, hangi yöntemin daha uygulanabilir, daha güvenli ve daha kolay olduğu belirlenmelidir. Bu amaçla üç DNA ekstraksiyonu protokolü kullanılmıştır. Bu protokollerden elde edilen DNA örneklerinin saflıkları ve konsantrasyonları karşılaştırılmıştır. Elde edilen DNA'nın kalitesi ve miktarı NanoDrop Spektrofotometre (A260 / 280 absorbans) ve agaroz jel elektroforezi ile belirlenmiştir. Ayrıca, ekstrakte edilen DNA'lar polimeraz zincir reaksiyonları (PCR) ile amplifiye edilmiş ve izole edilmiş DNA'ların  PCR reaksiyonları için uygunluğu da belirlenmiştir. Sonuçlar, CTAB DNA ekstraksiyon yöntemiyle diğer protokollere kıyasla buğdayda (Triticum avestivum L.) en yüksek DNA konsantrasyonlarının elde edildiğini göstermiştir.

Kaynakça

  • Abdel-Latif, A., & Osman, G. (2017). Comparison of three genomic DNA extraction methods to obtain high DNA quality from maize. Plant Methods, 13(1), 1.
  • Abouseadaa, H. H., Osman, G. H., Ramadan, A. M., Hassanein, S. E., Abdelsattar, M. T., Morsy, Y. B., & Gad, A. A. (2015). Development of transgenic wheat (Triticum aestivum L.) expressing avidin gene conferring resistance to stored product insects. BMC plant biology, 15(1), 183.
  • Aljanabi, S. M., & Martinez, I. (1997). Universal and rapid salt-extraction of high quality genomic DNA for PCR-based techniques. Nucleic acids research, 25(22), 4692-4693.Chai, J. F., Liu, X., & Jia, J. Z. (2006). A rapid isolation method of wheat DNA suitable for PCR analysis. Journal of Plant Genetic Resources, 7(2), 246-248.
  • Cristina, D., Ciuca, M., & Cornea, C. P. (2017). Comparison of four genomic DNA isolation methods from single dry seed of wheat, barley and rye. AgroLife Scientific Journal, 6(1), 84-91.
  • Dilworth, E., & Frey, J. E. (2000). A rapid method for high throughput DNA extraction from plant material for PCR amplification. Plant Molecular Biology Reporter, 18(1), 61-64.
  • Doyle J.J., & Doyle J. L. (1987). A rapid DNA isolation procedure for small quantities of fresh leaf tissue, Phytochem. Bull., 9, 11-15.
  • Kadry, A. K., Gamal, O., & Waeil, A. A. (2012). Genetic diversity and taxonomic relationships of some'Ipomoea'species based on analysis of RAPD-PCR and SDS-PAGE of seed proteins. Australian Journal of Crop Science, 6(6), 1088.
  • Khanuja, S. P., Shasany, A. K., Darokar, M. P., & Kumar, S. (1999). Rapid isolation of DNA from dry and fresh samples of plants producing large amounts of secondary metabolites and essential oils. Plant Molecular Biology Reporter, 17(1), 74-74.
  • Lee, P. Y., Costumbrado, J., Hsu, C. Y., Kim, Y. H. (2012). Agarose gel electrophoresis for the separation of DNA fragments. Journal of Visualized Experiments, (62), 3923. Moreira, P. A., & Oliveira, D. A. (2011). Leaf age affects the quality of DNA extracted from Dimorphandra mollis (Fabaceae), a tropical tree species from the Cerrado region of Brazil. Genetics and Molecular Research, 10(1), 353-358.
  • Murray, M. G., & Thompson, W. F. (1980). Rapid isolation of high molecular weight plant DNA. Nucleic acids research, 8(19), 4321-4326.
  • Paris, M., & Carter, M. (2000). Cereal DNA: a rapid high-throughput extraction method for marker assisted selection. Plant Molecular Biology Reporter, 18(4), 357-360.
  • Plaschke, J., Ganal, M. W., & Röder, M. S. (1995). Detection of genetic diversity in closely related bread wheat using microsatellite markers. Theoretical and Applied Genetics, 91(6-7), 1001-1007.
  • Pervaiz, Z. H., Turi, N. A., Khaliq, I., Rabbani, M. A., & Malik, S. A. (2011). Methodology: a modified method for high-quality DNA extraction for molecular analysis in cereal plants. Genetics and molecular research: GMR, 10(3), 1669-1673.
  • Röder, M. S., Plaschke, J., König, S. U., Börner, A., Sorrells, M. E., Tanksley, S. D., & Ganal, M. W. (1995). Abundance, variability and chromosomal location of microsatellites in wheat. Molecular and General Genetics MGG, 246(3), 327-333.
  • Sahu, S. K., Thangaraj, M., & Kathiresan, K. (2012). DNA extraction protocol for plants with high levels of secondary metabolites and polysaccharides without using liquid nitrogen and phenol. ISRN Molecular Biology, 2012.
  • Somers, D. J., Isaac, P., & Edwards, K. (2004). A high-density microsatellite consensus map for bread wheat (Triticum aestivum L.). Theoretical and Applied Genetics, 109(6), 1105-1114.
  • Sönmezoğlu, Ö. A., & Keskin, H. (2015). Determination of genetically modified corn and soy in processed food products. Journal of Applied Biology & Biotechnology, 3, 32-37.
  • Sönmezoğlu, Ö. A., & Terzi, B. (2018). Characterization of some bread wheat genotypes using molecular markers for drought tolerance. Physiology and molecular biology of plants, 24(1), 159-166.
  • Varma, A., Padh, H., & Shrivastava, N. (2007). Plant genomic DNA isolation: an art or a science. Biotechnology Journal: Healthcare Nutrition Technology, 2(3), 386-392.
  • Yadav, M. K., & Chand, P. (2018). Assessment of Genetic Diversity among Twenty Indian Wheat (Triticum aestivum L.) Cultivars using Simple Sequence Repeat (SSR) Markers. Int. J. Curr. Microbiol. App. Sci, 7(3), 1708-1717.
Toplam 20 adet kaynakça vardır.

Ayrıntılar

Birincil Dil İngilizce
Konular Mühendislik
Bölüm Makaleler
Yazarlar

Özlem Ateş Sönmezoğlu 0000-0002-3423-7095

Begüm Terzi Bu kişi benim 0000-0002-3264-2257

Yayımlanma Tarihi 31 Aralık 2019
Yayımlandığı Sayı Yıl 2019 Sayı: 17

Kaynak Göster

APA Ateş Sönmezoğlu, Ö., & Terzi, B. (2019). Comparison of DNA extraction protocols for PCR-based techniques in wheat. Avrupa Bilim Ve Teknoloji Dergisi(17), 860-865. https://doi.org/10.31590/ejosat.646491