title: FAST-FISH with laser beam microdissected DOP-PCR probe distinguishes the sex chromosomes of Silene latifolia.
abstract: We present an improved FISH strategy for differentiating the sex chromosomes of the dioecious model plant, Silene latifolia. Fixed mitotic protoplasts were dropped on a polyethylene naphthalate membrane, the X or Y chromosomes were isolated using nitrogen laser beam microdissection, catapulted by laser pressure, and amplified by DOP-PCR. A modified FAST-FISH protocol based on a short hybridization time combined with a low concentration of probe was used. The success of this approach is demonstrated by the differential labeling of the X and Y chromosomes and it could represent a quick method for comparing organization of plant genomes.
authors.affiliation: Institute of Biophysics, Academy of Sciences of the Czech Republic, Kralovopolska 135, CZ-612 65 Brno, Czech Republic.
authors.forename: Roman
authors.initials: R
authors.lastname: Hobza
authors.forename: Martina
authors.initials: M
authors.lastname: Lengerova
authors.forename: Halina
authors.initials: H
authors.lastname: Cernohorska
authors.forename: Jiri
authors.initials: J
authors.lastname: Rubes
authors.forename: Boris
authors.initials: B
authors.lastname: Vyskot
mesh_terms: D002232:Carbocyanines / Q000737:chemistry; D032461:Chromosomes, Plant / Q000235:genetics*; D015342:DNA Probes / Q000737:chemistry; D005456:Fluorescent Dyes / Q000737:chemistry; D017404:In Situ Hybridization, Fluorescence / Q000379:methods*; D007834:Lasers; D008677:Metaphase; D042282:Microdissection / Q000379:methods*; D016133:Polymerase Chain Reaction / Q000379:methods; D020401:Primed In Situ Labeling / Q000379:methods*; D011523:Protoplasts / Q000166:cytology; D012730:Sex Chromosomes / Q000235:genetics*; D029751:Silene / Q000235:genetics*
chemical_list: D002232:Carbocyanines; D015342:DNA Probes; D005456:Fluorescent Dyes; C089187:cyanine dye 3