Comparison of DNA Extraction Methods in Terms of Yield, Purity, Long-Term Storage, and Downstream Manipulation with Brewer's Yeast Chromosomal DNA

Abstract

Five methods of DNA isolation (a “classical” phenol/chloroform extraction and four commercially available kits) were compared in terms of the yield, purity, integrity, and stability of extracted DNA from both laboratory and industrial yeast strains. DNA isolation with commercial kits employing silica-based adsorption columns mostly gave lower yields of DNA. The integrity of DNA soon declines and after 6 months of storage under the manufacturer's instructions, it was almost fully degraded. Isolation of DNA using bead breaking of cells resulted in very low yield and also low purity of the DNA. The use of a commercial kit without silica columns and phenol/chloroform treatment as well as the traditional phenol/chloroform extraction were certainly more time consuming, but the extracted DNA was of high purity and concentration, with minimal damage, stable over months, and usable for subsequent molecular analyses (cloning, PCR, RFLP, DNA sequencing). Only one kit based on lyticase treatment, salting out without phenol/chloroform treatment, and “classical” phenol/chloroform extraction method allowed us to isolate a high yield of DNA with sufficient purity and stability in comparison with the use of commercial kits based on silica columns.