Prerequisite Protocols

• Ultra Fast TSS Competent E. coli Cell Prep or TSS Competent E. coli Cell Prep

Materials

• Intact Plasmid
• Chemically Competent E. Coli Cells
• Sterile 0.2mL PCR Tube
• Sterile 2mL Round Bottom Tube
• LB Broth (Miller Mod)
• LB Agar (Miller Mod) + Plasmid Antibiotics
• Sterile Petri Dishes
• Sterile Inoculation Loops

Equipment

• PCR Machine
• Ice Bucket
• Incubator Shaker
• Static 37C Incubator
• Small tray or box that can mount onto shaker incubator
• Pipettes (20uL, 10uL, 2uL)

Step By Step

1. Thaw TSS Competent Cells on ice. Preheat PCR machine to hold at 42C.