In situ RNA hybridization using Technovit resin in Arabidopsis thaliana

Katsuaki Takechi, Wataru Sakamoto, Maki Katsuhara, Minoru Murata, Fusao Motoyoshi

Research output: Contribution to journalArticle

8 Citations (Scopus)

Abstract

A protocol is described for RNA in situ hybridization using thin sections prepared by Technovit resin. Technovit is a widely used resin for histological examinations. Since it does not require time-consuming processes such as removal of the resin and can be performed without high temperature treatment, a high resolution of sections could be possible compared to other resins and paraffin. Thin sections (approximately 4 μm) were made from inflorescences of Arabidopsis thaliana embedded in Technovit 8100 resin, and in situ hybridization was performed using the protocol described in this article. Hybridization signals were observed using LEAFY and other genes as probes, showing that this resin can be used for in situ analysis. In our experiments, the most important factor for asuccessful in situ hybridization pattern was to optimize the RNase A concentration after hybridization. We routinely used RNase A at a concentration of 2-5 ng/ml, a concentration much lower than that used for paraffin embedding method. Thus, the use of the Technovit resin for plant tissue embedding results in a faster protocol and greater quality than allowed by paraffin sections.

Original languageEnglish
Pages (from-to)43-51
Number of pages9
JournalPlant Molecular Biology Reporter
Volume17
Issue number1
Publication statusPublished - 1999

Fingerprint

nucleic acid hybridization
Arabidopsis
resins
In Situ Hybridization
Pancreatic Ribonuclease
Arabidopsis thaliana
Resins
RNA
Paraffin
Plant Resins
Tissue Embedding
Paraffin Embedding
Inflorescence
alkanes
in situ hybridization
hybridization
Temperature
Genes
probes (equipment)
plant tissues

Keywords

  • Arabidopsis thaliana
  • In situ hybridization
  • LEAFY gene
  • Technovit resin

ASJC Scopus subject areas

  • Plant Science
  • Biochemistry, Genetics and Molecular Biology(all)
  • Biochemistry

Cite this

In situ RNA hybridization using Technovit resin in Arabidopsis thaliana. / Takechi, Katsuaki; Sakamoto, Wataru; Katsuhara, Maki; Murata, Minoru; Motoyoshi, Fusao.

In: Plant Molecular Biology Reporter, Vol. 17, No. 1, 1999, p. 43-51.

Research output: Contribution to journalArticle

Takechi, Katsuaki ; Sakamoto, Wataru ; Katsuhara, Maki ; Murata, Minoru ; Motoyoshi, Fusao. / In situ RNA hybridization using Technovit resin in Arabidopsis thaliana. In: Plant Molecular Biology Reporter. 1999 ; Vol. 17, No. 1. pp. 43-51.
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