Primer Design for PCR Amplification of Gene Flanks
2:49
Production of OSCAR Constructs
5:48
Agrobacterium tumefaciens Mediated Transformation (ATMT) of Fungi
7:45
Deletion Mutant Identification by PCR
8:53
Results: The OSCAR Method Generates a Gene Deletion Plasmid for Fungi in a Single Reaction
10:23
Conclusion
Transcript
The overall goal of this procedure is to generate precise gene deletion mutants in fungi. This method can help answer key questions in functional genetics of fungi, such as what role a particular gene of interest plays in the ability of a fungus t
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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.