Alpha 2.0
Login Register
» Articles » PMID: 21548894

Extraction of Genomic DNA from Yeasts for PCR-based Applications

Overview
Journal Biotechniques
Specialties Biomedical Engineering
Biotechnology
Date 2011 May 10
PMID 21548894
Citations 219
Authors
Marko Looke
Kersti Kristjuhan
Arnold Kristjuhan
Affiliations
Soon will be listed here.
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.

Citing Articles

PCR-fingerprinting of culturable yeasts from commercially obtained beers: a simple and engaging applied microbiological laboratory exercise.

Pfliegler W, Imre A, Biotechnology BSc Class Of U, Pocsi I MicroPubl Biol. 2025; 2025.

PMID: 40052139 PMC: 11883471. DOI: 10.17912/micropub.biology.001381.

Characterizing heterologous protein burden in Komagataella phaffii.

Kastberg L, Jacobsen I, Ozdemir E, Workman C, Jensen M, Forster J FEMS Yeast Res. 2025; 25.

PMID: 39971732 PMC: 11881926. DOI: 10.1093/femsyr/foaf007.

Cryptic genetic variation shapes the fate of gene duplicates in a protein interaction network.

Dibyachintan S, Dube A, Bradley D, Lemieux P, Dionne U, Landry C Nat Commun. 2025; 16(1):1530.

PMID: 39934115 PMC: 11814230. DOI: 10.1038/s41467-025-56597-0.

Challenges in elucidating ethylene glycol metabolism in Saccharomyces cerevisiae.

Senatore V, Masotti F, Milanesi R, Ceccarossi S, Maestroni L, Serra I FEMS Yeast Res. 2025; 25.

PMID: 39919757 PMC: 11878538. DOI: 10.1093/femsyr/foaf006.

Construction and iterative redesign of synXVI a 903 kb synthetic Saccharomyces cerevisiae chromosome.

Goold H, Kroukamp H, Erpf P, Zhao Y, Kelso P, Calame J Nat Commun. 2025; 16(1):841.

PMID: 39833175 PMC: 11747415. DOI: 10.1038/s41467-024-55318-3.

References
1.
Thompson J, Register E, Curotto J, Kurtz M, Kelly R . An improved protocol for the preparation of yeast cells for transformation by electroporation. Yeast. 1998; 14(6):565-71. DOI: 10.1002/(SICI)1097-0061(19980430)14:6<565::AID-YEA251>3.0.CO;2-B. View
2.
Akada R, Murakane T, Nishizawa Y . DNA extraction method for screening yeast clones by PCR. Biotechniques. 2000; 28(4):668-70, 672, 674. DOI: 10.2144/00284st02. View
3.
Sedman T, Joers P, Kuusk S, Sedman J . Helicase Hmi1 stimulates the synthesis of concatemeric mitochondrial DNA molecules in yeast Saccharomyces cerevisiae. Curr Genet. 2005; 47(4):213-22. DOI: 10.1007/s00294-005-0566-4. View
4.
Ito H, Fukuda Y, Murata K, Kimura A . Transformation of intact yeast cells treated with alkali cations. J Bacteriol. 1983; 153(1):163-8. PMC: 217353. DOI: 10.1128/jb.153.1.163-168.1983. View
5.
Ling M, Merante F, Robinson B . A rapid and reliable DNA preparation method for screening a large number of yeast clones by polymerase chain reaction. Nucleic Acids Res. 1995; 23(23):4924-5. PMC: 307485. DOI: 10.1093/nar/23.23.4924. View