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Rapid Deletion Production in Fungi via Agrobacterium Mediated Transformation of OSCAR Deletion Constructs
Chapters
- 00:05Title
- 01:02Primer Design for PCR Amplification of Gene Flanks
- 02:49Production of OSCAR Constructs
- 05:48Agrobacterium tumefaciens Mediated Transformation (ATMT) of Fungi
- 07:45Deletion Mutant Identification by PCR
- 08:53Results: The OSCAR Method Generates a Gene Deletion Plasmid for Fungi in a Single Reaction
- 10:23Conclusion
Gene deletion mutants generated through homologous recombination are the gold standard for gene function studies. The OSCAR (One Step Construction of Agrobacterium-Recombination-ready-plasmids) method for rapid generation of deletion constructs is described. Agrobacterium mediated fungal transformation follows. Finally, a PCR based confirmation method of gene deletions in fungal transformants is presented.
