FAST-FISH with laser beam microdissected DOP-PCR probe distinguishes the sex chromosomes of Silene latifolia.
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.
Hobza
Roman
R
Institute of Biophysics, Academy of Sciences of the Czech Republic, Kralovopolska 135, CZ-612 65 Brno, Czech Republic.
Lengerova
Martina
M
Cernohorska
Halina
H
Rubes
Jiri
J
Vyskot
Boris
B
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*
D002232:Carbocyanines; D015342:DNA Probes; D005456:Fluorescent Dyes; C089187:cyanine dye 3