This protocol is good for DNA extraction from microbial fungi isolated from aquatic insect guts. It works for small input tissues (starting from one fungal thallus). The product can be used for PCR and Sanger sequencing directly, which is ideal for fungal barcode analyses. This protocol is modified from a method described in Gottlieb and Lichtwardt 2001 (Mycologia Vol. 93, No. 1, pp. 66-81).