Počet záznamů: 1
Application of Flow Cytometry for Genome Size Determination in Geosmithia Fungi: A Comparison of Methods
- 1.
SYSNO ASEP 0436051 Druh ASEP J - Článek v odborném periodiku Zařazení RIV J - Článek v odborném periodiku Poddruh J Článek ve WOS Název Application of Flow Cytometry for Genome Size Determination in Geosmithia Fungi: A Comparison of Methods Tvůrce(i) Veselská, Tereza (MBU-M) ORCID
Svoboda, Jan (MBU-M) RID
Růžičková, Žaneta (MBU-M)
Kolařík, Miroslav (MBU-M) RID, ORCIDZdroj.dok. Cytometry. Part A. - : Wiley - ISSN 1552-4922
85A, č. 10 (2014), s. 854-861Poč.str. 8 s. Jazyk dok. eng - angličtina Země vyd. US - Spojené státy americké Klíč. slova fungi ; genome size ; flow cytometry Vědní obor RIV CE - Biochemie Institucionální podpora MBU-M - RVO:61388971 UT WOS 000342636200005 DOI https://doi.org/10.1002/cyto.a.22500 Anotace 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. Pracoviště Mikrobiologický ústav Kontakt Eliška Spurná, eliska.spurna@biomed.cas.cz, Tel.: 241 062 231 Rok sběru 2015
Počet záznamů: 1