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Description
  • Byly popsána nová metoda fluorescenční in situ hybridizace s vysokým rozlišením využívající jako templátu natažených mitotických chromozómů. Metoda umožňuje natažení mitotických chromozómů až na 100nasobek jejich původní délky. Protokol zahrnuje třídění chromozómů pomocí průtokové cytometrie, štěpení pomocí proteinázy K a natažení chromozómů směsí etanol:kyselina octová (3:1). Fluorescenční signály na natažených chromozómech jsou silnější než u standartních mitotických chromozómů a maximální citlivost dosahovala hodnoty 1kbp. Prostorové rozlišení dvou fluorescenčních signálů se podařilo zlepšit až na 70 kbp v porovnání se 5-10 Mbp u fluorescenční in situ hybridizace na mitotických chromozómech. (cs)
  • A novel high-resolution FISH (fluorescence in situ hybridisation) strategy using super stretching flow-sorted plant chromosomes as targets is described. The technique that allows longitudinal extension of chromosomes of more than 100-times its original metaphase size is especially attractive for FISH on plant species with large chromosomes, whose pachytene chromosomes are generally too long and heterochromatin patterns too complex for FISH analysis. The protocol involves flow cytometric sorting of metaphase chromosome, mild proteinase-K digestion of air-dried chromosomes on microscopic slides, followed by stretching with ethanol : acetic acid (3:1). Stretching ratios were assessed in a number of FISH experiments with super-stretched chromosomes from barley, wheat, rye and chickpea hybridised with 45S and 5S ribosomal DNAs, the [GAA]n microsatellite, the [TTTAGGG]n telomeric repeat, and a BAC clone as probes. FISH signals on stretched chromosomes were brighter than those on the untre...
  • A novel high-resolution FISH (fluorescence in situ hybridisation) strategy using super stretching flow-sorted plant chromosomes as targets is described. The technique that allows longitudinal extension of chromosomes of more than 100-times its original metaphase size is especially attractive for FISH on plant species with large chromosomes, whose pachytene chromosomes are generally too long and heterochromatin patterns too complex for FISH analysis. The protocol involves flow cytometric sorting of metaphase chromosome, mild proteinase-K digestion of air-dried chromosomes on microscopic slides, followed by stretching with ethanol : acetic acid (3:1). Stretching ratios were assessed in a number of FISH experiments with super-stretched chromosomes from barley, wheat, rye and chickpea hybridised with 45S and 5S ribosomal DNAs, the [GAA]n microsatellite, the [TTTAGGG]n telomeric repeat, and a BAC clone as probes. FISH signals on stretched chromosomes were brighter than those on the untre... (en)
Title
  • High-resolution FISH on super-stretched flow-sorted plant chromosomes
  • High-resolution FISH on super-stretched flow-sorted plant chromosomes (en)
  • FISH s vysokým rozlišením na natahovaných rostlinných chromozómech tříděných pomocí průtokové cytometrie (cs)
skos:prefLabel
  • High-resolution FISH on super-stretched flow-sorted plant chromosomes
  • High-resolution FISH on super-stretched flow-sorted plant chromosomes (en)
  • FISH s vysokým rozlišením na natahovaných rostlinných chromozómech tříděných pomocí průtokové cytometrie (cs)
skos:notation
  • RIV/61389030:_____/04:00109153!RIV/2005/AV0/A56005/N
http://linked.open.../vavai/riv/strany
  • 940;950
http://linked.open...avai/riv/aktivita
http://linked.open...avai/riv/aktivity
  • P(ME 527), P(ME 528), Z(AV0Z5038910)
http://linked.open...iv/cisloPeriodika
  • -
http://linked.open...vai/riv/dodaniDat
http://linked.open...aciTvurceVysledku
http://linked.open.../riv/druhVysledku
http://linked.open...iv/duvernostUdaju
http://linked.open...titaPredkladatele
http://linked.open...dnocenehoVysledku
  • 566177
http://linked.open...ai/riv/idVysledku
  • RIV/61389030:_____/04:00109153
http://linked.open...riv/jazykVysledku
http://linked.open.../riv/klicovaSlova
  • Chromosome streching; fluorescence in situ hybridization; chromosome sorting (en)
http://linked.open.../riv/klicoveSlovo
http://linked.open...odStatuVydavatele
  • GB - Spojené království Velké Británie a Severního Irska
http://linked.open...ontrolniKodProRIV
  • [13186619D691]
http://linked.open...i/riv/nazevZdroje
  • Plant Journal
http://linked.open...in/vavai/riv/obor
http://linked.open...ichTvurcuVysledku
http://linked.open...cetTvurcuVysledku
http://linked.open...vavai/riv/projekt
http://linked.open...UplatneniVysledku
http://linked.open...v/svazekPeriodika
  • 37
http://linked.open...iv/tvurceVysledku
  • Bartoš, Jan
  • Doležel, Jaroslav
  • Kubaláková, Marie
  • Valárik, Miroslav
  • Kovářová, Pavlína
  • de Jong, H.
http://linked.open...n/vavai/riv/zamer
issn
  • 0960-7412
number of pages
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