Rapid alkaline preparation for yeast circular covalently closed DNA molecules.
The alkaline preparation of prokaryotic plasmids (Birnboim and Doly, 1979) has been here adapted to yeast. By simple denaturation and renaturation steps we recovered, from Saccharomyces cerevisiae and Schizosaccharomyces pombe, a population of nucleic acid molecules highly enriched in circular forms. In S. cerevisiae killer strains it is possible to copurify double stranded RNA molecules. The overall recovery was estimated to be 10-30% of the total circular molecules.
D004270:DNA, Circular / Q000302:isolation & purification*; D004271:DNA, Fungal / Q000302:isolation & purification*; D004587:Electrophoresis, Agar Gel / Q000379:methods; D006863:Hydrogen-Ion Concentration; D009691:Nucleic Acid Denaturation; D009695:Nucleic Acid Renaturation; D010957:Plasmids; D012441:Saccharomyces cerevisiae / Q000235:genetics*; D004718:Saccharomycetales / Q000235:genetics*; D012568:Schizosaccharomyces / Q000235:genetics*
D004270:DNA, Circular; D004271:DNA, Fungal