Jones Lab:Protocols:Agrobacteria
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Competent Agrobacteria (for cold-shock transformation)
- O/N starter culture (with appropriate antibiotics; GV3101 should be grown in Rif and Gent), at 28°C
- Inoculate 500ml LB with starter culture
- Grow at 28°C until OD 0.5-1.0
- Chill culture on ice for 30 min
- Spin at 10k for 15 min in SS34 at 4°C
- Wash in 50 mL ice-cold 20mM CaCl2
- Spin at 10k for 15 min in SS34 at 4°C
- Resuspend in 10 mL ice-cold 20mM CaCl2
- 100 μL aliquots, flash-frozen in N2