Tyramide Signal Amplification: Fluorescence In Situ Hybridization for Identifying Homoeologous Chromosomes

Araceli Fominaya; Yolanda Loarce; Juan M González; Esther Ferrer

doi:10.1007/978-1-4939-3622-9_4

Tyramide Signal Amplification: Fluorescence In Situ Hybridization for Identifying Homoeologous Chromosomes

Methods Mol Biol. 2016:1429:35-48. doi: 10.1007/978-1-4939-3622-9_4.

Authors

Araceli Fominaya¹, Yolanda Loarce², Juan M González², Esther Ferrer²

Affiliations

¹ Department of Biomedicine and Biotechnology, University of Alcalá, Campus Universitario, Ctra Madrid-Barcelona Km 33,600, 28871, Alcalá de Henares, Madrid, Spain. araceli.fominaya@uah.es.
² Department of Biomedicine and Biotechnology, University of Alcalá, Campus Universitario, Ctra Madrid-Barcelona Km 33,600, 28871, Alcalá de Henares, Madrid, Spain.

PMID: 27511165
DOI: 10.1007/978-1-4939-3622-9_4

Abstract

Tyramide signal amplification (TSA) fluorescence in situ hybridization (FISH) has been shown as a valuable molecular tool for visualizing specific amplified DNA sequences in chromosome preparations. This chapter describes how to perform TSA-FISH, paying special interest to its two critical steps: probe generation and metaphase plate generation. The potential of physically mapping 12S-globulin sequences by TSA-FISH as a means of identifying homeology among chromosome regions of Avena species was tested and is discussed.

Keywords: Avena; Metaphase chromosomes; Obtaining probes; Physical mapping; TSA-FISH.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Amides / chemistry
Avena / genetics*
Chromosome Mapping / methods*
Chromosomes, Plant*
Fluorescent Dyes / chemistry
In Situ Hybridization, Fluorescence / methods*
Tyramine / chemistry

Substances

Amides
Fluorescent Dyes
Tyramine