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Molecular Cytogenetics

, 6:6

First Online: 04 February 2013Received: 21 November 2012Accepted: 14 December 2012

Abstract

BackgroundThe multicolor banding MCB-mBAND technique provides a unique opportunity to characterize intrachromosomal rearrangements and to determine chromosomal breakpoints. Until recently, MCB probes have only been available for human and some murine chromosomes. Generation of MCB probes for chromosomes of other species, useful and required in many cytogenetics research fields, was limited by technical difficulties. MCB probes are established by chromosome microdissection followed by whole genomic DNA amplification. However, unambiguous identification of the target chromosome is required for MCB-probe establishment. Previously proposed protocols suggested G-banding staining or preliminary FISH with whole chromosome paints WCP as methods to identify the chromosome of interest.

ResultsHere we present a complete workflow for MCB probe generation for those cases and species where chromosome morphology is too challenging to recognize target chromosomes by conventional methods and where WCP probes are not available. The workflow was successfully applied for murine chromosomes that are difficult to identify unambiguously. Additionally, we showed that glass-needle based microdissection enables establishment of a whole set of WCP paints by microdissection of individual chromosomes of a single metaphase

ConclusionsThe present method can be applied for generation of whole or region-specific DNA probes for species, where karyotyping of G-banded chromosomes is challenging due to similar chromosome morphology and-or chromosome banding patterns.

KeywordsChromosome painting FISH-banding Multicolor banding Chromosome microdissection AbbreviationsDNADeoxyribonucleic acid

PCRPolymerase chain reaction

DOP-PCRDegenerate oligonucleotide-primed PCR.

Electronic supplementary materialThe online version of this article doi:10.1186-1755-8166-6-6 contains supplementary material, which is available to authorized users.

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Autor: Nadezda Kosyakova - Ahmed Basheer Hamid - Arunrat Chaveerach - Krit Pinthong - Pornnarong Siripiyasing - Weerayuth Supiwong

Fuente: https://link.springer.com/







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