Comparison of different methods for extraction of mitochondrial DNA from human pathogenic yeasts.
Methods of rapidly extracting chromosomal DNA from human pathogenic yeasts were used in mitochondrial DNA (mtDNA) studies. This paper is concerned with rapid and reliable methods of extracting mtDNA for sequence analysis for species or strain identification, and epidemiological study of medically important fungi and fungal infections. To determine the optimal method of mtDNA extraction without isolating mitochondria, we examined three commonly used methods: 1). boiling, 2). glass bead disruption, and 3). a commercially available kit. We assessed the amount and quality of DNAs using a spectrophotometer and specific polymerase chain reaction (PCR). The DNA yield depended on the extraction method used and the yeast species. An adequate amount of mtDNA was obtained with both glass beads and a commercially available kit to amplify the mitochondrial gene using PCR without isolating the mitochondria. These techniques are convenient for extracting DNA from a variety of small-scale samples.
Yamada
Yohko
Y
Department of Chemical and Biological Sciences, Faculty of Science, Japan Women's University, Tokyo 112-8681, Japan. yohko@fc.jwu.ac.jp
Makimura
Koichi
K
Merhendi
Hossain
H
Ueda
Kumiko
K
Nishiyama
Yayoi
Y
Yamaguchi
Hideyo
H
Osumi
Masako
M
D017931:DNA Primers; D004272:DNA, Mitochondrial / Q000302:isolation & purification*; D006801:Humans; D066298:In Vitro Techniques; D003904:Mitosporic Fungi / Q000302:isolation & purification*; D016133:Polymerase Chain Reaction / Q000379:methods*
D017931:DNA Primers; D004272:DNA, Mitochondrial