IGEM:PennState/2006/Transformation

Transformation

Media/Reagents

• Competent cells
• SOC¹
• Antibiotic LB plates

Pre

• Add plasmid DNA to alloquats from electrocompetent cell prep
• Transfer DNA and Cells into electroporator cuvettes

Electroporation

1. Shock Cells, monitor discharge time
2. Add SOC to cuvettes (~400)
3. Transfer to culture tubes, shake at 37 for 1 hour (depending on antibiotic marker)
4. Plate ~200uL of SOC/Cells onto agar plate

¹To make SOC, add 20 μL 500 mM filter-sterilized glucose to 500 uL SOB (20 mM final glucose concentration)