title: An improved colony-PCR method for filamentous fungi for amplification of PCR-fragments of several kilobases.
abstract: A method was developed to perform PCR directly on mycelial pellets or colonies treated with NOVOzym 234. The method allows rapid screening of large numbers of transformants of both sporulating and non-sporulating fungi for the presence of (co)transforming plasmid copies or for specific genetic modifications such as gene disruption and site specific integration. PCR fragments of at least 3.2 kb can be obtained. Using this method the identification of specific disruption mutants from Aspergillus niger and Beauveria bassiana was carried out.
authors.affiliation: TNO Nutrition and Food Research Institute, Department of Molecular Genetics and Gene Technology, Zeist, The Netherlands.
authors.forename: C M
authors.initials: CM
authors.lastname: van Zeijl
authors.forename: E H
authors.initials: EH
authors.lastname: van de Kamp
authors.forename: P J
authors.initials: PJ
authors.lastname: Punt
authors.forename: G C
authors.initials: GC
authors.lastname: Selten
authors.forename: B
authors.initials: B
authors.lastname: Hauer
authors.forename: R F
authors.initials: RF
authors.lastname: van Gorcom
authors.forename: C A
authors.initials: CA
authors.lastname: van den Hondel
mesh_terms: D001234:Aspergillus niger / Q000201:enzymology / Q000235:genetics*; D003577:Cytochrome P-450 Enzyme System / Q000235:genetics; D004252:DNA Mutational Analysis / Q000379:methods*; D004271:DNA, Fungal / Q000235:genetics*; D004798:Enzymes / Q000494:pharmacology; D005087:Glucan 1,4-alpha-Glucosidase / Q000235:genetics; D010957:Plasmids / Q000235:genetics; D014170:Transformation, Genetic / Q000235:genetics
chemical_list: D004271:DNA, Fungal; D004798:Enzymes; C045154:novozym 234; D003577:Cytochrome P-450 Enzyme System; D005087:Glucan 1,4-alpha-Glucosidase