Prerequisite Protocols

• Agrobacterium Heat Shock Transformation
• Agrobacterium Colony PCR

Materials

• Flowering Petunia Plants (fresh blooms ideal but any will suffice)
• Agrobacterium tumefaciens GV3101 armed with plasmid and verified via PCR
• LBKGR Liquid Media
• LB Agar (Miller Mod)
• Agroinfiltration Buffer
• 1mL Oral Syringe (no needle, blunt tipped)
• 10uL Sterile Inoculation loop (or autoclaved toothpick)
• Sterile 50mL Centrifuge Tube

Equipment

• Orbital Shaker
• 250mL Erlenmeyer Flasks
• 28C Static Incubator
• Petri Dishes

Step By Step

1. Prepare LB Agar plates with Kan 100mg/L, Gent 100mg/L, and Rif 15mg/L.
2. Streak out your Agrobacterium cells, already transformed with your desired plasmid. Incubate at 28C overnight in the dark.