Plant Molecular Biology Reporter

, Volume 16, Issue 1, pp 90–90

A Rapid DNA Extraction Method for RFLP and PCR Analysis from a Single Dry Seed

  • Hee Wan Kang
  • Yong Gu Cho
  • Ung Han Yoon
  • Moo Young Eun
Article

DOI: 10.1023/A:1007418606098

Cite this article as:
Kang, H.W., Cho, Y.G., Yoon, U.H. et al. Plant Molecular Biology Reporter (1998) 16: 90. doi:10.1023/A:1007418606098

Abstract

A single-seed DNA extraction method was developed for rapid identification of plant genotype. The method was applied to 12 plant species, including the oil seeds sesame and soybean. The results were comparable to those obtained for oil-less seeds such as rice. This method will be useful for genotypic selection which requires rapid screening of large populations. It can also be used to identify varietal purity of seed stocks by PCR and RFLP analysis. The method includes two major steps, (i) treatment by proteinase K in an SDS extraction buffer, and (ii) grinding of a single half seed in the buffer after incubation. About 1.5–2 µg of DNA per half seed (the endosperm part) of rice was obtained and more than 200 half seed samples could be handled by one person in a day. The DNA could be used for fingerprinting and detection of target genes in a transgenic plant by PCR. The amplified PCR products from the half seed DNA exhibited the same banding patterns as those from leaf DNA. Yield and quality of DNA extracted from half seeds of rice was also sufficient for RFLP analysis. The remnant half seeds containing the embryo can be maintained for later germination of selected genotypes.

DNA extration DNA fingerprint half seed PCR RFLP target gene 

Copyright information

© Kluwer Academic Publishers 1998

Authors and Affiliations

  • Hee Wan Kang
    • 1
  • Yong Gu Cho
    • 1
  • Ung Han Yoon
    • 1
  • Moo Young Eun
    • 1
  1. 1.National Institute of Agricultural Science & TechnologySuwonKorea

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