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Current issue: Volume 22, No. 2 (2022)
Umbraško I., Batjuka A., Petjukevičs A., Škute N.. 2022. Evaluation of the effectiveness of using a universal method for isolating genomic dsDNA by salting out technique according to the S.M. Aljanabi and I. Martinez protocol for yeast Saccharomyces cerevisiae. Acta Biol. Univ. Daugavp., 22 (2): 195 – 202.
Abstract
This work evaluated the possibility of using the universal salting out genomic dsDNA isolation method according to the S.M. Aljanabi and I. Martinez protocol from yeast cells of Saccharomyces cerevisiae also for possible PCR-based applications. Our modification of this protocol
adapted it for the extraction of DNA from the liquid culture of S. cerevisiae yeast colonies/ cells. This protocol did not require any hazardous chemicals or specific conditions and was specially developed for the simultaneous processing of many samples. Prepared suspension liquid of 100 µL according to the McFarland Standard method corresponds to 1.5×108 cells per volume unit. Turbidity standards provide the opportunity to visually determine the number of cells per unit volume by turbidity. For fungi, a culture with a value of 0.5 McFarland Standard contains 30 times less microbial cells than enterobacteria. Approximately 400 ng/µL of total genomic dsDNA (S. cerevisiae) can be isolated from 1.5×108 S. cerevisiae cells. The isolated dsDNA had good quality and purity (A260/A280 ~1.8-2.0 ±0.2-0.3 and A260/A230 in the range of 2.0-2.2 ±0.2-0.3). Isolated dsDNA corresponds to high molecular weight dsDNA with high levels of nucleic acids in 1 µL of solution. This modified DNA extraction technique is as a rapid and universal low-cost genomic dsDNA isolation protocol that is suitable for yeast cell liquid cultures.
Keywords: DNA concentration, DNA electrophoresis, DNA extraction, Saccharomyces
cerevisiae.
*Corresponding author: Inta Umbraško. Daugavpils University, Institute of Life Sciences and Technology, Parades Str. 1A, Daugavpils, Latvia, LV-5401, E-mail: inta.umbrasko@du.lv
Anna Batjuka. Daugavpils University, Institute of Life Sciences and Technology, Parades Str.
1A, Daugavpils, Latvia, LV-5401, Email: aleksandrs.petjukevics@du.lv Aleksandrs Petjukevičs. Daugavpils University, Institute of Life Sciences and Technology, Parades Str. 1A, Daugavpils, Latvia, LV-5401, Email: aleksandrs.petjukevics@du.lv Nataļja Škute. Daugavpils University, Institute of Life Sciences and Technology, Parades Str. 1A, Daugavpils, Latvia, LV-5401, Email: natalja.skute@du.lv
