Felicia Zlati, Andreas Müsgens, Lars M Blank, Nick Wierckx, Kerstin Schipper
Reliable genomic DNA isolation is essential for investigating the genetic basis of any organism. In fungi such as Ustilago maydis, this task is particularly challenging due to the rigid cell wall and the presence of polysaccharides, pigments, and other compounds that can compromise DNA quality as contaminants. This chapter provides practical guidance on overcoming these challenges by detailing robust cell lysis strategies, including enzymatic digestion and mechanical disruption, which form the critical first step in gDNA extraction. Additionally, a set of protocols is presented, tailored to different downstream applications, from PCR amplification to high-throughput short- and long-read sequencing for complex genome assemblies.