title: Extraction of genomic DNA from yeasts for PCR-based applications.
abstract: We have developed a quick and low-cost genomic DNA extraction protocol from yeast cells for PCR-based applications. This method does not require any enzymes, hazardous chemicals, or extreme temperatures, and is especially powerful for simultaneous analysis of a large number of samples. DNA can be efficiently extracted from different yeast species (Kluyveromyces lactis, Hansenula polymorpha, Schizosaccharomyces pombe, Candida albicans, Pichia pastoris, and Saccharomyces cerevisiae). The protocol involves lysis of yeast colonies or cells from liquid culture in a lithium acetate (LiOAc)-SDS solution and subsequent precipitation of DNA with ethanol. Approximately 100 nanograms of total genomic DNA can be extracted from 1 × 10(7) cells. DNA extracted by this method is suitable for a variety of PCR-based applications (including colony PCR, real-time qPCR, and DNA sequencing) for amplification of DNA fragments of ≤ 3500 bp.
authors.affiliation: Institute of Molecular and Cell Biology, University of Tartu, Tartu, Estonia.
authors.forename: Marko
authors.initials: M
authors.lastname: Lõoke
authors.forename: Kersti
authors.initials: K
authors.lastname: Kristjuhan
authors.forename: Arnold
authors.initials: A
authors.lastname: Kristjuhan
mesh_terms: D000085:Acetates / Q000032:analysis* / Q000737:chemistry*; D002176:Candida albicans / Q000235:genetics; D004271:DNA, Fungal / Q000302:isolation & purification* / Q000378:metabolism*; D007716:Kluyveromyces / Q000235:genetics; D010843:Pichia / Q000235:genetics*; D016133:Polymerase Chain Reaction / Q000379:methods; D012441:Saccharomyces cerevisiae / Q000235:genetics; D012568:Schizosaccharomyces / Q000235:genetics; D012967:Sodium Dodecyl Sulfate / Q000032:analysis* / Q000737:chemistry*; D015003:Yeasts / Q000235:genetics
chemical_list: D000085:Acetates; D004271:DNA, Fungal; D012967:Sodium Dodecyl Sulfate; C488804:lithium acetate