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(Reference retrieved automatically from Web of Science through information on FAPESP grant and its corresponding number as mentioned in the publication by the authors.)

Universal fluorescence in situ hybridization (FISH) protocol for mapping repetitive DNAs in insects and other arthropods

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Author(s):
Cabral-de-Mello, Diogo Cavalcanti [1, 2] ; Marec, Frantisek [2]
Total Authors: 2
Affiliation:
[1] UNESP Univ Estadual Paulista, Inst Biociencias, Dept Biol Geral & Aplicada, BR-13506900 Rio Claro, SP - Brazil
[2] Czech Acad Sci, Biol Ctr, Inst Entomol, Ceske Budejovice - Czech Republic
Total Affiliations: 2
Document type: Journal article
Source: Molecular Genetics and Genomics; v. 296, n. 3 FEB 2021.
Web of Science Citations: 1
Abstract

Repetitive DNAs comprise large portion of eukaryote genomes. In genome projects, the assembly of repetitive DNAs is challenging due to the similarity between repeats, which generate ambiguities for alignment. Fluorescence in situ hybridization (FISH) is a powerful technique for the physical mapping of various sequences on chromosomes. This technique is thus very helpful in chromosome-based genome assemblies, providing information on the fine architecture of genomes and their evolution. However, various protocols are currently used for FISH mapping, most of which are relatively laborious and expensive, or work properly only with a specific type of probes or sequences, and there is a need for a universal and affordable FISH protocol. Here we tested a FISH protocol for mapping of different DNA repeats, such as multigene families (rDNAs, U snDNAs, histone genes), satellite DNAs, microsatellites, transposable elements, DOP-PCR products, and telomeric motif (TTAGG)(n), on the chromosomes of various insects and other arthropods. Different cell types and stages obtained from diverse tissues were used. The FISH procedure proved high quality and reliable results in all experiments performed. We obtained data on the chromosomal distribution of DNA repeats in representatives of insects and other arthropods. Thus, our results allow us to conclude that the protocol is universal and requires only time adjustment for chromosome/DNA denaturation. The use of this FISH protocol will facilitate studies focused on understanding the evolution and role of repetitive DNA in arthropod genomes. (AU)

FAPESP's process: 19/19069-7 - Investigating the evolution of satellite DNAs in holocentric chromosomes of pest species
Grantee:Diogo Cavalcanti Cabral de Mello
Support Opportunities: Regular Research Grants