About: Application of Flow Cytometry for Genome Size Determination in Geosmithia Fungi: A Comparison of Methods

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Attributes	Values
rdf:type	skos:Concept http://linked.opendata.cz/ontology/domain/vavai/Vysledek
rdfs:seeAlso	http://dx.doi.org/10.1002/cyto.a.22500
Description	Genome size has played an important role in the evolution of plants and animals because changes in genome size seem to accompany if not facilitate evolutionary adaptation to environmental conditions. Flow cytometry (FCM) is a widespread method for determining genome size thanks to its high accuracy and speed of measurements. Nevertheless, only a few comparative studies of FCM methods exist in the field of mycology, and reviews are absent. In this study, we compared the suitability of several concentrations and RNAse A incubation times, fixatives and buffers for estimating genome size in fungi. We chose the genus Geosmithia as a model filamentous fungus. We also introduced a new standard, Aspergillus fumigatus CEA10, to determine absolute genome size. We found FCM to be an appropriate method for measuring genome size in fungi, but optimization steps showed that incorrect propidium iodide staining of nuclei can overestimate genome size due to cytoplasmic staining. We identified fixation with methanol:glacial acetic acid (3:1 v/v), 10% DMSO, 0.1% Triton-X 100, and 5 mM EDTA in combination with Tris-MgCl2 buffer as the best treatment. (c) 2014 International Society for Advancement of Cytometry Genome size has played an important role in the evolution of plants and animals because changes in genome size seem to accompany if not facilitate evolutionary adaptation to environmental conditions. Flow cytometry (FCM) is a widespread method for determining genome size thanks to its high accuracy and speed of measurements. Nevertheless, only a few comparative studies of FCM methods exist in the field of mycology, and reviews are absent. In this study, we compared the suitability of several concentrations and RNAse A incubation times, fixatives and buffers for estimating genome size in fungi. We chose the genus Geosmithia as a model filamentous fungus. We also introduced a new standard, Aspergillus fumigatus CEA10, to determine absolute genome size. We found FCM to be an appropriate method for measuring genome size in fungi, but optimization steps showed that incorrect propidium iodide staining of nuclei can overestimate genome size due to cytoplasmic staining. We identified fixation with methanol:glacial acetic acid (3:1 v/v), 10% DMSO, 0.1% Triton-X 100, and 5 mM EDTA in combination with Tris-MgCl2 buffer as the best treatment. (c) 2014 International Society for Advancement of Cytometry (en)
Title	Application of Flow Cytometry for Genome Size Determination in Geosmithia Fungi: A Comparison of Methods Application of Flow Cytometry for Genome Size Determination in Geosmithia Fungi: A Comparison of Methods (en)
skos:prefLabel	Application of Flow Cytometry for Genome Size Determination in Geosmithia Fungi: A Comparison of Methods Application of Flow Cytometry for Genome Size Determination in Geosmithia Fungi: A Comparison of Methods (en)
skos:notation	RIV/00216208:11310/14:10289248!RIV15-MSM-11310___
http://linked.open...avai/riv/aktivita	I S
http://linked.open...avai/riv/aktivity	I, S
http://linked.open...iv/cisloPeriodika	10
http://linked.open...vai/riv/dodaniDat	2015
http://linked.open...aciTvurceVysledku	Veselská, Tereza Kolařík, Miroslav
http://linked.open.../riv/druhVysledku	J - Článek v odborném periodiku
http://linked.open...iv/duvernostUdaju	S - Úplné a pravdivé údaje nepodléhající ochraně podle zvláštních právních předpisů
http://linked.open...titaPredkladatele	Univerzita Karlova v Praze / Přírodovědecká fakulta
http://linked.open...dnocenehoVysledku	3734
http://linked.open...ai/riv/idVysledku	RIV/00216208:11310/14:10289248
http://linked.open...riv/jazykVysledku	eng - angličtina
http://linked.open.../riv/klicovaSlova	flow cytometry; genome size; fungi (en)
http://linked.open.../riv/klicoveSlovo	fungi genome size flow cytometry
http://linked.open...odStatuVydavatele	US - Spojené státy americké
http://linked.open...ontrolniKodProRIV	[1F2F9A5770AB]
http://linked.open...i/riv/nazevZdroje	Cytometry Part A
http://linked.open...in/vavai/riv/obor	CE
http://linked.open...ichTvurcuVysledku	2 (xsd:int)
http://linked.open...cetTvurcuVysledku	4 (xsd:int)
http://linked.open...UplatneniVysledku	2014
http://linked.open...v/svazekPeriodika	85A
http://linked.open...iv/tvurceVysledku	Svoboda, Jan Kolařík, Miroslav Růžičková, Žaneta Veselská, Tereza
http://linked.open...ain/vavai/riv/wos	000342636200005
issn	1552-4922
number of pages	8 (xsd:int)
http://bibframe.org/vocab/doi	10.1002/cyto.a.22500
http://localhost/t...ganizacniJednotka	11310

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