Combined Fluorescent-Chromogenic In Situ Hybridization for Identification and Laser Microdissection of Interphase Chromosomes

• Autores: Paz, Nerea; Zabala, Amaia; Royo, Félix; García-Orad, África.; Zugaza, José Luis; Parada, Luis Antonio
• Año de publicación: 2013
• Idioma: inglés
• Tipo de recurso: artículo
• Materia: Interphase chromosomes; Laser microdissection; FISH-CISH; Down syndrome
• Institución: Consejo Nacional de Investigaciones Científicas y Técnicas

Chromosome territories constitute the most conspicuous feature of nuclear architecture, and they exhibit non-random distribution patterns in the interphase nucleus. We observed that in cell nuclei from humans with Down Syndrome two chromosomes 21 frequently localize proximal to one another and distant from the third chromosome. To systematically investigate whether the proximally positioned chromosomes were always the same in all cells, we developed an approach consisting of sequential FISH and CISH combined with laser-microdissection of chromosomes from the interphase nucleus and followed by subsequent chromosome identification by microsatellite allele genotyping. This approach identified proximally positioned chromosomes from cultured cells, and the analysis showed that the identity of the chromosomes proximally positioned varies. However, the data suggest that there may be a tendency of the same chromosomes to be positioned close to each other in the interphase nucleus of trisomic cells. The protocol described here represents a powerful new method for genome analysis
Fil: Paz, Nerea.
Fil: Zabala, Amaia.
Fil: Royo, Félix.
Fil: García-Orad, África..
Fil: Zugaza, José Luis.
Fil: Parada Lopez, Luis Antonio. Consejo Nacional de Invest.cientif.y Tecnicas. Centro Cientifico Tecnol.conicet - Salta. Instituto de Patologia Experimental;