An optimized method for high-quality DNA extraction medicinal fungi Mycoleptodonoides aitchisonii for whole genome sequencing

Abstract

Abstract The extraction of pure high molecular weight DNA and collecting genomic DNA from fungi is difficult. This is because filamentous fungi has a sturdy cell wall, high protein and polysaccharide levels resistant to the usual DNA extraction procedures. A low-cost and reliable DNA extraction method was designed from Mycoleptodonoides. aitchisonii fit for whole-genome sequencing for identification and mapping of the A mating-type gene. Mycoleptodonoides. aitchisonii belongs to the Climacodontaceae family has pharmaceutical activities. In the present study, the mycelia of M. aitchisonii, which grew from the different concentrations of malt extract and minimal liquid media, have been compared systematically to determine the DNA extraction procedure resulted in DNA concentration with excellent purity and quality. The best protocol that resulted in good quality DNA was further validated using polymerase chain reaction amplification with a specific primer to amplify the homeodomain protein (Mahd2-18) gene that encodes a transcription factor. The method proposed DNA extraction using CTAB (Cetyl Trimethyl Ammonium Bromide) method and purify by commercial kit from mycelium grown in the minimal liquid media give the best result with the high concentration of DNA for whole-genome sequencing by Next Generation Sequencing and other applications.