Open AccessExtraction of genomic DNA from yeasts for PCR-based applications
Author(s) -
Marko Lõoke,
Kersti Kristjuhan,
Arnold Kristjuhan
Publication year - 2017
Publication title -
biotechniques
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.617
H-Index - 131
eISSN - 1940-9818
pISSN - 0736-6205
DOI - 10.2144/000114497
Subject(s) - genomic dna , schizosaccharomyces pombe , kluyveromyces lactis , biology , yeast , dna , dna extraction , candida parapsilosis , polymerase chain reaction , candida albicans , lysis , ethanol precipitation , genomic library , saccharomyces cerevisiae , multiple displacement amplification , microbiology and biotechnology , biochemistry , gene , ethanol , base sequence
Protocol Summary 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 × 107 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.
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