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Ultra-simple DNA extraction method for marker-assisted selection using microsatellite markers in rice

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Abstract

We prevent an ultra-simple DNA extraction method for microsatellite analysis of rice. Each extraction requires only one microtube, one disposable pipette tip, TE buffer and few pieces (about 5 mm) of rice leaf tissue. This is sufficient for 200 PCR reactions. The extract can be kept in the freezer for long-term storage. Also, DNA can be extracted from 200–300 individuals in a few hours. These features enabled us to perform rapid largescale seedling genotyping required for marker-assisted selection. We have also examined the applicability of this method for other PCR-based markers: RAPDs, nuclear STS, chloroplast STS and chloroplast microsatellites.

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Abbreviations

QTL:

quantitative trait locus

RAPD:

random amplified polymorphic DNA

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Authors and Affiliations

  1. Graduate School of Science and Technology, Kobe University, Nada-ku, 657-8501, Kobe, Japan

    Nobuyuki Ikeda & Nonnatus S. Bautista

  2. National Institute of Agrobiological Resources, 305-8602, Kannondai, Tsukuba, Japan

    Tetsuya Yamada

  3. Faculty of Agriculture, Kobe University, Nada-ku, 657-8501, Kobe, Japan

    Osamu Kamijima & Takashige Ishii

Authors
  1. Nobuyuki Ikeda
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  2. Nonnatus S. Bautista
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  3. Tetsuya Yamada
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  4. Osamu Kamijima
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  5. Takashige Ishii
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Corresponding author

Correspondence to Takashige Ishii.

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Ikeda, N., Bautista, N.S., Yamada, T. et al. Ultra-simple DNA extraction method for marker-assisted selection using microsatellite markers in rice. Plant Mol Biol Rep 19, 27–32 (2001). https://doi.org/10.1007/BF02824075

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  • DOI: https://doi.org/10.1007/BF02824075

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