Fluorescence In Situ Hybridization and Rehybridization Using Bacterial Artificial Chromosome Probes

Elzbieta Stankiewicz; Tianyu Guo; Xueying Mao; Yong-Jie Lu

doi:10.1007/978-1-4939-9769-5_16

Fluorescence In Situ Hybridization and Rehybridization Using Bacterial Artificial Chromosome Probes

Methods Mol Biol. 2019:2054:243-261. doi: 10.1007/978-1-4939-9769-5_16.

Authors

Elzbieta Stankiewicz¹, Tianyu Guo¹, Xueying Mao¹, Yong-Jie Lu²

Affiliations

¹ Centre for Molecular Oncology, Barts Cancer Institute, Queen Mary University of London, London, UK.
² Centre for Molecular Oncology, Barts Cancer Institute, Queen Mary University of London, London, UK. y.j.lu@qmul.ac.uk.

PMID: 31482460
DOI: 10.1007/978-1-4939-9769-5_16

Abstract

Fluorescence in situ hybridization (FISH) method enables in situ genetic analysis of both metaphase and interphase cells from different types of material, including cell lines, cell smears, and fresh and paraffin-embedded tissue. Despite the growing number of commercially available FISH probes, still for large number of gene loci or chromosomal regions commercial probes are not available. Here we describe a simple method for generating FISH probes using bacterial artificial chromosomes (BAC). Due to genome-wide coverage of BAC clones, there are almost unlimited possibilities for the analysis of any genomic regions using BAC FISH probes.

Keywords: Bacterial artificial chromosome; Fluorescence in situ hybridization; Probes; Rehybridization; Tissue pretreatment.

MeSH terms

Bacteriological Techniques / instrumentation
Bacteriological Techniques / methods
Cell Culture Techniques / methods
Cell Line
Chromosomes, Artificial, Bacterial / genetics*
DNA Probes / genetics
DNA Probes / isolation & purification*
DNA, Bacterial / genetics
DNA, Bacterial / isolation & purification*
Deoxyuracil Nucleotides / chemistry
Dideoxynucleotides / chemistry
Digoxigenin / analogs & derivatives
Digoxigenin / chemistry
Fluoresceins / chemistry
Fluorescent Dyes / chemistry
Frozen Sections
Genomics / instrumentation
Genomics / methods*
Humans
In Situ Hybridization, Fluorescence / instrumentation
In Situ Hybridization, Fluorescence / methods*
Rhodamines / chemistry
Staining and Labeling / instrumentation
Staining and Labeling / methods
Uridine Triphosphate / analogs & derivatives
Uridine Triphosphate / chemistry

Substances

DNA Probes
DNA, Bacterial
Deoxyuracil Nucleotides
Dideoxynucleotides
Fluoresceins
Fluorescent Dyes
Rhodamines
digoxigenin-11-deoxyuridine triphosphate
fluorescein-12-dideoxyuridine triphosphate
tetramethylrhodamine
Digoxigenin
Uridine Triphosphate