Camparison of Extraction Techniques for Isolation Fungal DNA

Abstract

For effective molecular screening of fungi using polymerase chain reaction, a simple, quick, reliable, and inexpensive DNA extraction technique is always obliged. Because of the low sensitivity of culture-based detection methods, PCRbased techniques have been developed to amplify DNA from fungal infections. In PCR-based assay, pure DNA must be required, as well as an easy-to-follow DNA extraction approach. Due to amount of fungus in a tissue biopsy or clinical sample may be relatively small, extraction efficiency is extremely crucial in PCR-based medical diagnostic applications. In this study, we are comparing four different methods of DNA extraction and compared their efficacy in the extraction of DNA from filamentous fungi commonly isolated in our laboratory. It was found that the effectiveness of cell wall disruption decreasing in the following order: GeNei™ Fungal Genomic DNA Extraction Teaching Kit > SDS containing extraction buffer > CTAB containing extraction buffer and vortex with glass beads > CTAB and SDS containing extraction buffer. Keywords: DNA isolation, DNA extraction methods, Filamentous fungi, Polymerase chain reaction