Fluorescence in situ hybridization in oat

Abstract

This chapter describes methods to detect gene loci or gene transcripts by fluorescence labeling. Fluorescence in situ hybridization (FISH) can be used to identify the positions of genes or BACs or the distribution of repetitive sequences on metaphase chromosomes as well as the identification of alien chromosomes. It enables the identification of gene loci and active transcription sites in interphase nuclei and also the localization of cellular transcripts. The protocols here deal with the production of DNA and RNA probes, the preparation of oat metaphase spreads and root tissue sections, the subsequent hybridization, post-­hybridization washes, and detection by immunofluorescence.