Abstract
Fluorescent in situ hybridization coupled with immunofluorescence (FISH/IF) is an assay that has been widely used to study DNA-protein interactions. The technique is based on the use of a fluorescent nucleic acid probe and fluorescent antibodies to reveal the localization of a DNA sequence and a specific protein in the cell. The interaction can be inferred by the quantification of the co-localization between the protein and the DNA. Here, we describe a detailed FISH/IF methodology that our group used to study RPA-telomere interaction in the pathogenic protozoa parasite Trypanosoma cruzi.
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Acknowledgments
The authors want to thank Simone Calderano for helping with the photos presented in Fig. 1. M.C.E. is a fellow from the National Council for Scientific and Technological Development (CNPq: 306199/2018-1). RSP received a Fundação de Amparo à Pesquisa do Estado de São Paulo fellowship (FAPESP 2014/02978-0).
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Pavani, R.S., Elias, M.C. (2021). Following Trypanosoma cruzi RPA-DNA Interaction Using Fluorescent In Situ Hybridization Coupled with Immunofluorescence (FISH/IF). In: Oliveira, M.T. (eds) Single Stranded DNA Binding Proteins. Methods in Molecular Biology, vol 2281. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-1290-3_12
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DOI: https://doi.org/10.1007/978-1-0716-1290-3_12
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