Abstract
Publisher Summary This chapter describes a set of simple DNA-related experiments that have used to characterize basic genomic and genetic properties of the ligninolytic basidiomycete Phanerochaete chrysosporium . The set consists of (1) rapid DNA preparation and construction of genomic libraries; (2) genome size estimation by dot-blot hybridization; (3) use of CsCl-bisbenzimide gradients to separate mitochondrial, ribosomal, and chromosomal DNA and to estimate percentage gas chromatography; (4) investigation of DNA sequence variation between different strains and use of restriction fragment length polymorphisms (RFLPs) for spore analysis and genetic mapping of cloned sequences. The chapter describes a rapid method for isolation of DNA from filamentous fungi. The procedure starts from freeze-dried material and gives high yields of high-molecular-weight DNA suitable for restriction and ligation. It can be used for processing of many samples in parallel in microfuge tubes. Except for the RNase treatment, all steps are done at room temperature.
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