A rapid and simple method for the preparation of yeast mitochondrial DNA.

A rapid method for the extraction of yeast mitochondrial DNA (mtDNA) is described. The method is based on: preparation of spheroplastis, lysis of the cellular membrane, isopicnic purification of mitochondria, lysis of the mitochondrial membranes, and mtDNA isolation without using CsCl gradient centrifugation. In comparison with previous methods (1, 2), the method described here consists of rapid and simple-to-perform steps, and does not require neither phenol treatment nor the use of carcinogenic dyes (ethidium bromide and bis-benzimide). The resulting mtDNA is sufficiently pure for RFLP analyses, and the yield is high even from wild strains.