An improved colony-PCR method for filamentous fungi for amplification of pcr-fragments of several kilobases
article
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.
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.
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.
Topics
Colony-PCR methodFilamentous fungiRapid screeningFungiColony-PCR methodFilamentous fungiBiotechnologyAspergillus nigerBacterium transformationFungusGene disruptionGenetic transformationMyceliumNonhumanPlasmidPolymerase chain reactionPriority journalAspergillus nigerCytochrome P-450 Enzyme SystemDNA Mutational AnalysisDNA, FungalEnzymesGlucan 1,4-alpha-GlucosidasePlasmidsTransformation, GeneticAspergillus nigerBacteria (microorganisms)BassianaBeauveriaCordyceps bassianaFungi
TNO Identifier
234333
ISSN
01681656
Source
Journal of Biotechnology, 59(3), pp. 221-224.
Place of publication
Amsterdam, Netherlands
Pages
221-224
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