Agrobacterium transformation problems - Trouble preparing agrobacterium (Oct/24/2008 )

Hi all,

Am currently trying to insert a plasmid into agrobacterium. The plasmid has amp resistance and the agrobacterium line carb and rfp resistance. Am using LB agar plates with rfp and amp at various concentrations and incubating the plates overnight at 28 degree's. When i check the plates the next morning the whole plate is covered in agrobacterium and it is impossible to isolate indivdual colonies. In theroy the plates should take 2 days for colonies to appear. Ive tried increasing the antibiotic levels and also reducing incubation time and starting plasmid concentration but this is no use! Can anyone tell me what im doing wrong????????

Thanks!