Total DNA preparation excluding extracellular acidic polysaccharide from Lipomyces yeasts and its application to taxonomic studies.
A preparation method of total DNA from Lipomyces yeasts was improved in order to exclude extracellular acidic polysaccharide thoroughly. The method combined an ultracentrifuge and polyethylene glycol precipitation with the usual method. The total DNAs obtained were analyzed for G + C content and by DNA-DNA hybridization. The results all agreed almost completely with literature data. All the DNA samples prepared using this method were pure enough for these taxonomic analyses and could also be used as templates of PCR for amplification of small subunit ribosomal DNA and the internal transcribed spacer region.
Nishimura
Katsushi
K
Department of Materials and Applied Chemistry, College of Science and Technology, Nihon University, Tokyo, Japan. nisimura@chem.cst.nihon-u.ac.jp
Shimada
Koji
K
Iwasawa
Hiroki
H
Naganuma
Takafumi
T
Uzuka
Yasuyuki
Y
D002965:Classification; D004271:DNA, Fungal / Q000302:isolation & purification*; D009693:Nucleic Acid Hybridization; D011092:Polyethylene Glycols / Q000737:chemistry; D011134:Polysaccharides / Q000737:chemistry*; D012331:RNA, Fungal / Q000096:biosynthesis / Q000302:isolation & purification; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014461:Ultracentrifugation; D015003:Yeasts / Q000737:chemistry* / Q000145:classification*
D004271:DNA, Fungal; D011134:Polysaccharides; D012331:RNA, Fungal; D011092:Polyethylene Glycols