magistrsko delo
Darja Kolar (Author), Jasna Dolenc Koce (Reviewer), Lilijana Bizjak-Mali (Mentor), Lilijana Bizjak-Mali (Thesis defence commission member), Aleš Kladnik (Thesis defence commission member), Jasna Dolenc Koce (Thesis defence commission member), Rok Kostanjšek (Thesis defence commission member), Aleš Kladnik (Co-mentor)

Abstract

Namen raziskave je bil ugotoviti, ali so krvne celice proteusov (Proteus anguinus) iz arhivske zbirke primeren material za oceno velikosti jedrnega genoma s pomočjo slikovne citometrije. Krvne celice smo vzorčili iz ventrikla srca arhiviranih živali in barvali po Feulgenu s Schiffovim reagentom, ki specifično barva DNA. Meritve integrirane optične gostote (IOG) so bile izvedene na jedrih rdečih krvničk. Kot standard smo uporabili krvne celice arhiviranih osebkov nektura (Necturus maculosus), kot kontrolo pa krvne razmaze, pripravljene iz sveže krvi. Izračunane vrednosti C so bile zelo variabilne in so večinoma močno odstopale od znane ocenjene velikosti genoma proteusa v bazi Animal Genome Size Database. Krvne celice, izolirane iz srca živali iz arhivske zbirke, so bile neuporabne za meritve IOG in izračun vrednosti C, saj so bile živali večinoma neprimerno fiksirane in arhivirane. Vrednosti C primerljive objavljenim podatkom, smo dobili na vzorcih svežih krvnih razmazov in izoliranih krvnih celicah osebkov, fiksiranih bodisi v etanolu bodisi v formaldehidu, ki so bili nato arhivirani v etanolu. Za nadaljnje raziskave velikosti genoma s slikovno citometrijo so najbolj primerni sveži krvni razmazi, slednje velja tudi za nektura kot standard.

Keywords

evropska jamska repata dvoživka;endemit Dinarskega krasa;velikost genoma;vrednost C;slikovna citometrija;

Data

Language: Slovenian
Year of publishing:
Typology: 2.09 - Master's Thesis
Organization: UL BF - Biotechnical Faculty
Publisher: [D. Kolar]
UDC: 597.92(043.2)
COBISS: 5331023 Link will open in a new window
Views: 789
Downloads: 185
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Other data

Secondary language: English
Secondary title: Genome size analyses in population of proteus with image analysis cytometry
Secondary abstract: The aim of the study was to determine, whether the proteus (Proteus anguinus) blood cells from the archive collection provide a suitable material for estimation of nuclear genome size by image cytometry. Blood cells were sampled from the ventricle of the heart of archived animals and stained by Feulgen reaction with a Schiff reagent, which specifically stains DNA. Integrated optical density (IOD) was measured in red blood cell nuclei. The blood cells of archived specimens of common mudpuppy (Necturus maculosus) were used as standard, and blood smears prepared from fresh blood were used as control. The calculated C-values were highly variable and mostly differ from the known size of the proteus genome in the Animal Genome Size Database. Blood cells isolated from the hearts of animals from the archive were not suitable for IOD measurements and C-value calculations, since animals were mostly improperly fixed and archived. C-values comparable to published data were obtained on fresh blood smear samples and blood cells isolated from specimens fixed either in ethanol or formaldehyde and subsequently archived in ethanol. For further genome size studies with image cytometry, fresh blood smears are the most suitable. This also applies to common mudpuppy as standard.
Secondary keywords: European cave salamander;endemic for Dinaric karst;genome size;C-value;image analysis cytometry;
Type (COBISS): Master's thesis/paper
Study programme: 0
Embargo end date (OpenAIRE): 1970-01-01
Thesis comment: Univ. Ljubljana, Biotehniška fak.
Pages: VIII, 50, [1] f.
ID: 11221903