Visualization of Polytene Chromatin in Mosquito Cell Nuclei Using Three-Dimensional Fluorescence In Situ Hybridization
- 1Department of Entomology, Virginia Polytechnic Institute and State University, Blacksburg, Virginia 24060, USA
- 2Department of Genetics and Cell Biology, Tomsk State University, Tomsk 634050, Russia
- 3Laboratory of Evolutionary Genomics of Insects, Institute of Cytology and Genetics, Siberian Branch of the Russian Academy of Sciences, Novosibirsk 630090, Russia
- ↵4Correspondence: igor{at}vt.edu
Abstract
Chromosomes are intricately folded within the cell nucleus and interact with peripheral nuclear proteins. The chromatin architecture has a profound effect on how the genome is organized. 3D-FISH is a powerful technique that can reveal the structural and functional organization of chromosomes in the nuclear space. Here, we present a protocol for visualizing specific genomic regions in whole-mount paraformaldehyde-fixed cell nuclei of Anopheles mosquitoes. This protocol was tested in our laboratories and has been showed to be effective and reliable for visualizing genomic regions of various lengths—from 1-kb gene-scale fragments to chromosome-scale segments of DNA.
Footnotes
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From the Mosquitoes collection, edited by Laura B. Duvall and Benjamin J. Matthews.
