Publication details

Comparison of DNA extraction methods in terms of yield, purity, long-term storage and downstream manipulation with brewer's yeast chromosomal DNA



Year of publication 2014
Type Article in Periodical
Magazine / Source The Journal of the American Society of Brewing Chemists
MU Faculty or unit

Faculty of Science

Field Microbiology, virology
Keywords brewer's yeast; isolation of DNA; phenol/chloroform extraction; purification
Description 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 collumns mostly gave lower yields of DNA, integrity of DNA soon declines and after six months of storage under the manufacturer's instructions the DNA was almost fully degraded. Isolation of DNA by using bead breaking of cells resulted in very low yield and low purity of the DNA also. The use of a commercial kit without silica collumns 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 to isolate high yield of DNA with sufficient purity and stability in comparison with the use of commercial kits based on silica columns.
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