A modified procedure for isolation of yeast mitochondrial DNA.
A modified, rapid and inexpensive method for preparation of mitochondrial DNA (mtDNA), suitable for molecular analysis is proposed. It comprises batch cultivation of Saccharomyces cerevisiae strain NBIMCC 583 on a simple nutrient medium at 28 degrees C; permeabialization of cells from late exponential growth phase with cetyltrimethylamonnium bromide, mechanical disintegration of the cell wall; preparation of a mitochondrial fraction and subsequent isolation and purification of mtDNA. The amount and the purity of the obtained mtDNA have been checked and its application for molecular analysis proven. The main advantages of the proposed procedure for isolation of mtDNA are introduction of simple nutrient medium, replacement of the enzymatic lysis of the cell wall by the cheaper mechanical one, avoidance of ultracentrifugation steps and use of harmful chemical substances.
Nedeva
Trayana
T
Department of General and Industrial Microbiology, Faculty of Biology, The Sofia University St. Kliment Ohridski, Bulgaria. nedeva@biofac.uni-sofia.bg
Petrova
Ventzislava
V
Hristozova
Tsonka
T
Kujumdzieva
Anna
A
D004271:DNA, Fungal / Q000235:genetics / Q000302:isolation & purification*; D004272:DNA, Mitochondrial / Q000235:genetics / Q000302:isolation & purification*; D007202:Indicators and Reagents; D011110:Polymorphism, Genetic; D012150:Polymorphism, Restriction Fragment Length; D012441:Saccharomyces cerevisiae / Q000737:chemistry* / Q000235:genetics
D004271:DNA, Fungal; D004272:DNA, Mitochondrial; D007202:Indicators and Reagents