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Rapid and simple methodology for isolation of high quality genomic DNA from coniferous tissues (Taxus baccata)

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Abstract

Various investigations have been so far performed for extraction of genomic DNA from plant tissues, in which the extracted intact DNA can be exploited for a diverse range of biological studies. Extraction of high quality DNA from leathery plant tissues (e.g., coniferous organs) appears to be a critical stage. Moreover, for some species such as Taxus trees, bioprocess engineering and biosynthesis of secondary metabolites (e.g., paclitaxel) is a crucial step due to the restrictions associated with extinction of these species. However, extraction of intact genomic DNA from these plants still demands a rapid, easy and efficient protocol. To pursue such aim, in the current work, we report on the development of a simple and highly efficient method for the extraction of DNA from Taxus baccata. Based upon our protocol, interfering phenolic compounds were removed from extraction using polyvinylpyrrolidone and RNA contamination was resolved using LiCl. By employing this method, high quality genomic DNA was successfully extracted from leaves of T. baccata. The quality of extracted DNA was validated by various techniques such as RAPD marker, restriction digestions and pre-AFLP. Upon our findings, we propose this simple method to be considered for extraction of DNA from leathery plant tissues.

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Acknowledgments

The authors wish to express their gratitude to Ali Hamedbarghi and Fatemeh Zununi for their kind assistance.

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Authors and Affiliations

  1. Agricultural Biotechnology Research Institute for Northwest & West of Iran, Tabriz, Iran

    Abolfazl Barzegari

  2. Faculty of Biology, Tarbiat Moallem Azarbayjan University, Tabriz, Iran

    Sepideh Zununi Vahed

  3. Research Center for Pharmaceutical Nanotechnology, Faculty of Pharmacy, Tabriz University of Medical Sciences, Tabriz, Iran

    Sina Atashpaz, Sajjad Khani & Yadollah Omidi

  4. School of Advanced Biomedical Sciences, Tabriz University of Medical Sciences, Tabriz, Iran

    Sina Atashpaz, Sajjad Khani & Yadollah Omidi

Authors
  1. Abolfazl Barzegari
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  2. Sepideh Zununi Vahed
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  3. Sina Atashpaz
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  4. Sajjad Khani
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  5. Yadollah Omidi
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Corresponding author

Correspondence to Yadollah Omidi.

Additional information

A. Barzegari, S. Z. Vahed, S. Atashpaz contributed equally to this work and should be considered as co-first authors.

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Barzegari, A., Vahed, S.Z., Atashpaz, S. et al. Rapid and simple methodology for isolation of high quality genomic DNA from coniferous tissues (Taxus baccata). Mol Biol Rep 37, 833–837 (2010). https://doi.org/10.1007/s11033-009-9634-z

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  • DOI: https://doi.org/10.1007/s11033-009-9634-z

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