Leaf-punch method to prepare a large number of PCR templates from plants for SNP analysis

Sachiko Shiokai, Hiroyasu Kitashiba, Kenta Shirasawa, Kuniaki Nagano, Takeshi Nishio

Research output: Contribution to journalArticle

5 Citations (Scopus)

Abstract

DNA preparation is indispensable for genotyping by DNA polymorphism analysis, and that for a large number of plants is laborious. In the present study, a small leaf disk of rice, 1-2 mm in diameter, punched by a mini cork borer was found to be directly usable as a PCR template. DNA fragments <300 bp were amplified efficiently. Leaf disks of 1-1.5 mm in diameter were better than those of 2 mm for a small volume of reaction mixture. Multiplex PCR was possible with four or eight primer pairs using the small leaf disk as a template. Leaf disks of Arabidopsis, Lotus, wheat, soybean, tomato, Chinese cabbage, and melon were also good PCR templates. This method for preparation of PCR templates, named the leaf-punch method, was applicable to SNP analysis of a large number of plants by dot-blot-SNP analysis.

Original languageEnglish
Pages (from-to)329-336
Number of pages8
JournalMolecular Breeding
Volume23
Issue number2
DOIs
Publication statusPublished - 2009 Feb 1

Keywords

  • DNA preparation
  • Large-scale genotyping
  • Marker-assisted selection
  • PCR template
  • SNP analysis

ASJC Scopus subject areas

  • Biotechnology
  • Molecular Biology
  • Agronomy and Crop Science
  • Genetics
  • Plant Science

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