Prerequisite Protocols

• Columnless Plasmid Miniprep
• TSS Competent Agrobacterium Cell Prep

Materials

• Miniprepped Agrobacterium Plasmid (100ng/uL ideal)
• Agrobacterium tumefaciens GV3101, made competent via TSS Competent Agrobacterium Cell Prep
• Sterile 0.2mL PCR Tube
• Sterile 2mL Round Bottom Tube
• LB Broth (Miller Mod)
• LBKGR Solid Media
• Sterile Petri Dishes
• Sterile Inoculation Loops

Equipment

• PCR Machine set to hold at 37C
• Ice Bucket
• Tube Thermoshaker set to 28C 1000rpm (or room temp orbital shaker)
• Static 28C Incubator
• Pipettes (1000uL, 100uL, 10uL)

Step By Step

1. Thaw TSS Competent Cells on ice. Preheat PCR machine to hold at 37C.