A rapid, high yield mini-prep method for isolation of total genomic DNA from fungi.

Fungal genetic studies require a rapid method of isolating DNA from a large number of samples for restriction enzyme analysis. Creative Commons License This work is licensed under a Creative Commons Attribution-Share Alike 4.0 License. This regular paper is available in Fungal Genetics Reports: http://newprairiepress.org/fgr/vol35/iss1/11 Complementation studies including mts(MN9), cpc-1 (CD86 and j5) and mts(MN1) were performed as already described for mts MNl) (Koch and Bartheless, 1987). The mutants were recessive to their respective wild type alleles, but complementation of the amino acid analogue sensitive phenotype was not observed in heterocaryons carrying mutant alleles simultaneously. These findings suggest that cpc-1, mts(MN9) as well as mts(MNl) belong to the same complementation group. Institut für Angewandte Genetik, Universität Hannover, 3000 Hannover FRG. Supported by the Deutsche Forschungsgemeinschaft. Lee, S.B., M.G. Milgroom # Fungal genetic studies require a rapid method of isolating DNA from a large number of and J.W. Taylor samples for restriction enzyme analysis. Previous methods we have used are limited by relaA rapid, high yield mini-prep method tively low yield of 50 ug DNA/0.1g lyophilized mycelium (Zolan, M.E. and P.J. Pukkila 1986. Mol. for isolation of total genomic DNA Cell. Biol. 6:195-200) or tedious gel exclusion column chromatography (Biel, S.W. and F.W. from fungi. Parrish 1986. Anal. Biochem. 154:21-25). In addition, these two methods yielded no readily digestable DNA from Phytophthora cinnamomi. The following method facilitates rapid isolation of large quantities of easily digested total, genomic DNA from several species of Phytophthora, including P. cinnamomi, and several species of Boletus, Chroogomphus vinicolor, Gomphidius glutinosus, Leccinum manzanitae, Magnaporthe grisea, Neurospora crassa, N. tetrasperma, Omphalotus olivascens, and Talaromyces flavus. isolation of DNA from two Yield was increased to 200 ug DNA/0.1 g lyophilized mycelium and to three times as many samples can be achieved using this rapid method (current record is 64 isolates in one day versus 24 using previous methods). DNA has been successfully cut with all restriction enzymes tried to date. Solutions needed: 1. Lysis buffer: 50 mM Tris-HCl 50 mM EDTA 3% SDS 1% 2-mercaptoethanol (add just before use) 2. Chloroform:phenol (1:1) 3. SEVAG (chloroform:isoamyl alcohol, 24:1) 4. 3 M NaOAc (pH 8.0) 5. Isopropanol 6. Ethanol (100%, -20°C)