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An improved colony-PCR method for filamentous fungi for amplification of pcr-fragments of several kilobases

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Author: Zeijl, C.M.J. van · Kamp, E.H.M. van de · Punt, P.J. · Selten, G.C.M. · Hauer, B. · Gorcom, R.F.M. van · Hondel, C.A.M.J.J. van den
Type:article
Date:1998
Place: Amsterdam, Netherlands
Institution: Centraal Instituut voor Voedingsonderzoek TNO
Source:Journal of Biotechnology, 3, 59, 221-224
Identifier: 234333
doi: DOI:10.1016/S0168-1656(97)00170-3
Keywords: Nutrition · Colony-PCR method · Filamentous fungi · Rapid screening · Fungi · Colony-PCR method · Filamentous fungi · Biotechnology · Aspergillus niger · Bacterium transformation · Fungus · Gene disruption · Genetic transformation · Mycelium · Nonhuman · Plasmid · Polymerase chain reaction · Priority journal · Aspergillus niger · Cytochrome P-450 Enzyme System · DNA Mutational Analysis · DNA, Fungal · Enzymes · Glucan 1,4-alpha-Glucosidase · Plasmids · Transformation, Genetic · Aspergillus niger · Bacteria (microorganisms) · Bassiana · Beauveria · Cordyceps bassiana · Fungi

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.