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An improved method to extract DNA from mango Mangifera indica

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Abstract

High quality genomic DNA is the first step in the development of DNA-based markers for fingerprinting and genetic diversity of crops, including mango (Mangifera indica L.), a woody perennial. Poor quality genomic DNA hinders the successful application of analytical DNA-based tools. Standard protocols for DNA extraction are not suitable for mango since the extracted genomic DNA often contains secondary metabolites that interfere with analytical applications. In this study, we employed an additional step to remove polysaccharides, polyphenols and secondary metabolites from genomic DNA extracted from young or mature leaf tissue; then a modified traditional cetyl trimethyl ammonium bromide (CTAB) method was applied. The use of 0.4 M glucose improved DNA quality and avoided contamination and browning by polyphenolics, relative to the traditional CTAB method. This is an easy and efficient method for genomic DNA extraction from both young and mature leaves of mango. The isolated DNA was free of polysaccharides, polyphenols, RNA and other major contaminants, as judged by its clear colour, its viscosity, A260/A280 ratio and suitability for PCR-based reactions. This modified protocol was also used to extract high quality genomic DNA from other woody perennials, including walnut, guava, lychee, pear, grape and sugarcane.

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Abbreviations

CTAB:

cetyl trimethyl ammonium bromide

EDTA:

ethylenediamine tetraacetic acid

ISSR:

inter simple sequence repeat

PVP:

polyvinylpyrrolidone

TAE:

Tris acetate-EDTA

TE:

Tris-EDTA

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Authors and Affiliations

  1. Biotechnology Research Institute, Chinese Academy of Agricultural Sciences, Beijing, 100081, People’s Republic of China

    Mohammad S. Uddin, Wenli Sun & Qi Cheng

  2. Bangladesh Agricultural Research Institute, Gazipur, 1701, Bangladesh

    Mohammad S. Uddin

  3. College of Agriculture, Guangxi University, Nanning, 530004, People’s Republic of China

    Xinhua He

  4. Miki-cho post office, P. O. Box 7, Ikenobe 3011-2, Kagawa-ken, 761-079, Japan

    Jaime A. Teixeira da Silva

Authors
  1. Mohammad S. Uddin
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  2. Wenli Sun
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  3. Xinhua He
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  4. Jaime A. Teixeira da Silva
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  5. Qi Cheng
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Corresponding author

Correspondence to Qi Cheng.

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Electronic supplementary material. The online version of this article (DOI:10.2478/s11756-013-0311-2) contains supplementary material, which is available to authorized users.

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Uddin, M.S., Sun, W., He, X. et al. An improved method to extract DNA from mango Mangifera indica . Biologia 69, 133–138 (2014). https://doi.org/10.2478/s11756-013-0311-2

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