Identification of Positive GATEWAY Expression Clones Protocol Protocol for the identification of positive GATEWAY expression clones when both the pENTRY and pDEST vectors contain the same marker for bacterial selection. Protocol describes ways in which difficult vector combinations can be used effectively to obtain the appropriate expression clone without having to convert the pENTRY clone or pDEST clone to vectors with compatible antibiotic resistances.
PCR Analysis of Agrobacterium Protocol Protocol describes a method to determine the presence of plasmid DNA in an Agrobacterium culture. Compared to selection of transformed Agrobacterium, which can be ambiguous and normally takes several days for resistant colonies to appear, the approach described here is both rapid and accurate.
Transposon-Mediated Mutagenesis Protocol Protocol for transposon-mediated mutagenesis. Includes: Amplify ORF from MG1655; Transposition reaction; Transformation; Picking; Verify mutation by Culture PCR; Confirm mutations by sequencing; Curing the temperature sensitive pKD46 plasmid.