DNA isolation from Aspergillus flavus: Optimal method selection

Abstract

The methods for fungal genomic DNA isolation for PCR amplification, including commercially available kits, must often be adapted in order to produce sufficient amounts of high-quality DNA from specific fungal species. The aim of this study was to select an optimal method for the isolation of DNA from Aspergillus flavus suitable for PCR reaction. Four different methods were compared according to their efficiency in isolating pure DNA, their price and time consumption. DNA quantification and purity estimation were performed using the NanoDropTM 1000 UV/VIS spectrophotometer and DNA integrity and PCR products were determined by gel-electrophoresis. DNA quantity ranged from 92.77 ± 11.52 to 5477.4 ± 22.03 ng/µL, with A260/280 from 1.14 ± 0.10 to 1.94 ± 0.16, and A260/230 0.37 ± 0.05 to 1.91 ± 0.17. There were also great differences in time consumption per sample, ranging from 1 hr 15 min to 7 hr 5 min. The determined costs per sample were ranging from 0.12 € to 2.29 € per sample. All tested methods were suitable for the isolation of A. flavus genomic DNA and subsequently for PCR reaction.