Physical Genome Mapping Using Fluorescence In Situ Hybridization with Mosquito Chromosomes
The development of genomic resources and tools is an important step in designing novel approaches to genetic control of mosquitoes. Physical genome maps enhance the quality of the genome assemblies, improve gene annotation, and provide a better framework for comparative and population genomics studies in mosquitoes. In this chapter, we describe protocols for an important procedure in physical genome mapping—fluorescence in situ hybridization (FISH). We provide details on (1) dissection of salivary glands, ovaries, and imaginal discs for obtaining high-quality polytene or mitotic chromosome preparations; (2) DNA-labeling procedures and extraction of repetitive DNA fractions; and (3) approaches to FISH on polytene and mitotic chromosomes.
Key wordsMosquitoes Fluorescence in situ hybridization Polytene and mitotic chromosomes Physical genome mapping
Physical mapping of Anopheles was supported by Russian Science Foundation 15-14-20011 to Igor V. Sharakhov and physical mapping of Aedes and Culex was supported the National Institutes of Health grants 1R21AI121853 and 1R21AI123967 to Maria V. Sharakhova. We thank Phillip George for providing images of ovary dissection and Melissa Wade for proofreading the text.
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