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  • Physical Genome Mapping Using Fluorescence In Situ Hybridization with Mosquito Chromosomes.

Physical Genome Mapping Using Fluorescence In Situ Hybridization with Mosquito Chromosomes.

Methods in molecular biology (Clifton, N.J.) (2018-11-11)
Maria V Sharakhova, Gleb N Artemov, Vladimir A Timoshevskiy, Igor V Sharakhov
ABSTRACT

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.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Bovine Serum Albumin, heat shock fraction, protease free, pH 7, ≥98%
Sigma-Aldrich
Dextran sulfate sodium salt from Leuconostoc spp., for molecular biology, average Mw >500,000 (dextran starting material), contains 0.5-2% phosphate buffer