Automation protocol for high-efficiency and high-quality genomic DNA extraction from Saccharomyces cerevisiae

Abstract

Although many protocols have been previously developed for genomic DNA (gDNA) extraction from S. cerevisiae, to take advantage of recent advances in laboratory automation and DNA-barcode sequencing, there is a need for automated methods that can provide high-quality gDNA at high efficiency. Here, we describe and demonstrate a fully automated protocol that includes five basic steps: cell wall and RNA digestion, cell lysis, DNA binding to magnetic beads, washing with ethanol, and elution. Our protocol avoids the use of hazardous reagents (e.g., phenol, chloroform), glass beads for mechanical cell disruption, or incubation of samples at 100°C (i.e., boiling). We show that our protocol can extract gDNA with high efficiency both from cells grown in liquid culture and from colonies grown on agar plates. We also show results from gel electrophoresis that demonstrate that the resulting gDNA is of high quality.

Antibacterial Activity of Yellow Pigment from Micrococcus flavus AK 11 Isolates of the Coral Porites sp. from Gosong Beach, Central Java, Indonesia

“…In line with this, Alperovich et al (2023) stated that a good DNA purity is close to 1.8-2.0, and the use of the kit method can produce a DNA with fairly high purity. After that, amplification was carried out by the PCR and followed by electrophoresis of 1% agarose and obtained a product with a size of 1500bp (Alperovich et al, 2023). The 16S rRNA gene sequences of strains AK 11 contained 1437bp.…”