Abstract
Fluorescence in situ hybridization (FISH) is a widely used technique to detect and localize specific DNA or RNA sequences in cells. Although supplanted in many ways by fluorescently labeled DNA binding proteins, FISH remains the only cytological method to examine many genetic loci at once (up to six), and can be performed in any cell type and genotype. These advantages have proved invaluable in studying the spatial relationships between chromosome regions and the dynamics of chromosome segregation in bacteria. A detailed protocol for DNA FISH in E. coli is described.
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Acknowledgments
We thank Beth Weiner, Nancy Kleckner, Sota Hiraga, and Lucy Shapiro for sharing their detailed FISH protocols, which formed the basis of our methods. We thank Jeff Carmichael (Chroma Technology Corp) for technical assistance with fluorescent filters, and Po J. Chen and Anna K. Barker for comments on the manuscript. All work was supported by NIH Grant GM102679 to D.B.
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Visser, B.J., Joshi, M.C., Bates, D. (2017). Multilocus Imaging of the E. coli Chromosome by Fluorescent In Situ Hybridization. In: Espéli, O. (eds) The Bacterial Nucleoid. Methods in Molecular Biology, vol 1624. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7098-8_16
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DOI: https://doi.org/10.1007/978-1-4939-7098-8_16
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