Team:Groningen/Protocols for Lactococcus

From 2010.igem.org

Preparation of cells

• 10 ml ON culture (SMGG) in 100 ml SMGG
• Grow to OD600=0.2-0.7
• Wash three times with 50 ml icecold wash buffer
• Resuspend in 1 ml wash buffer

Electroporation

• 1 μl DNA in 40 μl cell-suspension in ice cold cuvette
• Electroporate at 2.5 kV, 25 μF, 200 Ohm.
• Add 4 ml SMG17MC
• Incubate 2 hours (Ery induction=50 ng/ml)
• Concentrate cells to 0.5 ml.
• Plate on GSM17- Agar.