Rapid purification of yeast mitochondrial DNA in high yield.
A procedure is presented for the rapid isolation of mitochondrial DNA (mtDNA) in high yield from Saccharomyces cerevisiae. Yeast cells, which may be grown to late stationary phase, are broken by a combination of enzymatic and mechanical means; mtDNA is then isolated from a crude mitochondrial lysate by a single cycle of bisbenzimide-CsCl buoyant density centrifugation. mtDNA so isolated is at least 99.5% pure, and has a mean duplex molecular weight of 24.5 . 10(6). In addition to mtDNA and bulk nuclear DNA, several other yeast nucleic acid species, identified as ribosomal DNA and a mixture of duplex RNAs, form discrete bands in these gradients.
D002458:Cell Fractionation; D002499:Centrifugation, Density Gradient; D002586:Cesium; D002712:Chlorides; D004271:DNA, Fungal / Q000302:isolation & purification*; D004272:DNA, Mitochondrial / Q000302:isolation & purification*; D008970:Molecular Weight; D012441:Saccharomyces cerevisiae / Q000032:analysis*
D002712:Chlorides; D004271:DNA, Fungal; D004272:DNA, Mitochondrial; D002586:Cesium; C028019:cesium chloride