Abstract

OSORIO CADAVID, Esteban; RAMIREZ, Mauricio; LOPEZ, William Andrés  and  MAMBUSCAY, Luz Adriana. Standarization of a simple protocol for yeast genomic DNA extraction. Rev. colomb. biotecnol [online]. 2009, vol.11, n.1, pp.125-131. ISSN 0123-3475.

A quick, simple and low-cost protocol for yeast DNA extraction by enzymatic treatment and alcoholic precipitations for cell wall lysis was standardized. By using Beta-glucuronidase instead of Zymolyase higher yields of DNA were obtained (124.9±30.2 ng/µl) with very high quality (A260/A280 nm = 1.86±0.1) and suitable for molecular biology assays. Moreover, a DNAse inactivation step by incubation at 100° C, was also introduced to ensure further DNA stability. DNA quality was assayed by PCR amplification of the ITS1-5.8S-ITS2 region, showing defined, quantifiable bands between 380 and 880 bp. These results are ideal for molecular identification and phylogenetic studies.

Keywords : Yeast; DNA extraction; Beta-Glucoronidase.

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