title: Localization of Low-Copy DNA Sequences on Mitotic Chromosomes by FISH.
abstract: Fluorescence in situ hybridization (FISH) is a widely used method to localize DNA sequences on mitotic and meiotic chromosomes and interphase nuclei. It was developed in early 1980s and since then it has contributed to numerous studies and important discoveries. Over the decades, the protocol was modified for ease of use, allowing for localizing multiple probes simultaneously and increasing its sensitivity and specificity. Despite the continuous improvements, the ability to detect short single-copy sequences of only a few kilobases or less, such as genes, remains limited. Here, we provide a detailed protocol for detection of short, single- or low-copy sequences on plant mitotic metaphase chromosomes.
authors: Centre of the Region Haná for Biotechnological and Agricultural Research, Institute of Experimental Botany, Šlechtitelů 31, CZ-78374, Olomouc, Czech Republic. karafiatova@ueb.cas.cz.
authors: Miroslava
authors: M
authors: Karafiátová
authors: Centre of the Region Haná for Biotechnological and Agricultural Research, Institute of Experimental Botany, Šlechtitelů 31, CZ-78374, Olomouc, Czech Republic.
authors: Jan
authors: J
authors: Bartoš
authors: Centre of the Region Haná for Biotechnological and Agricultural Research, Institute of Experimental Botany, Šlechtitelů 31, CZ-78374, Olomouc, Czech Republic.
authors: Jaroslav
authors: J
authors: Doležel
mesh_terms: D002874:Chromosome Mapping / Q000379:methods*; D032461:Chromosomes, Plant; D056915:DNA Copy Number Variations; D018744:DNA, Plant / Q000235:genetics*; D001467:Hordeum / Q000235:genetics*; D017404:In Situ Hybridization, Fluorescence / Q000379:methods*; D008938:Mitosis
chemical_list: D018744:DNA, Plant
keywords: Cell cycle synchronization; Cytogenetic mapping; Fluorochrome; Low-copy probe; Mitotic metaphase chromosomes; Post-fixation; Probe purification; Quantum yield