Lactobacilli are of considerable technological and commercial importance because of their role in the manufacturing and preservation of many fermented food products. The aim of this study was to optimize and evaluate three methods of DNA extraction and purification of DNA from Lactobacillus spp to be used for the amplification of 584-bp region of the tuf gene by polymerase chain reaction. The methods were: Phenol–chloroform extraction, boiling, and Wizard genomic DNA purification kit with modifications. Results demonstrated that extracted genomic DNA using Wizard genomic DNA purification kit with modifications for DNA extraction provided was higher yield of DNA with the highest purity than the other extraction methods. Purity was documented by gel electrophoresis. The quality of the genomic DNA isolated by this method was verified by polymerase chain reaction targeting the tuf gene. Keywords: Lactobacillus, DNA extraction, identification, tuf gene, PCR
