Cell density: Plate the cells at low density in order to avoid contacts at the beginning of the treatment. Split the cells at 1:30 when confluent.
Antibiotics: Several antibiotics have been tested in the lab. For now the most efficient has been ciprofloxin.
Surfactin: - Stock solution 1000x at 40mM in 100% ethanol (in the cold room)
- Use at 40M in the appropriate medium. Maintain the cells in culture for 2 weeks
Cipro: - Stock solution 1000x at 10mg/ml in water (centrifuge to collect debris after resuspension and filter the supernatant). Aliquots in 15ml tubes are stored at -20oC
- Use at 10g/ml in the appropriate medium. Maintain the cells in culture for 2 weeks
BM-Cyclin: - As recommended by Boehringer Ref: 799050 (Ref: 14879900 on the box)
Resuspension with 10ml water at 2.5mg/ml for BM-Cyclin 1
1.25mg/ml for BM-Cyclin 2
Aliquots of 400ul are stored at -20oC
- Use BM-Cyclin 1 at 10g/ml and BM-Cyclin 2 at 5g/ml
Cultivate the cells for 3 days in BM-Cyclin 1 containing medium
remove the medium and cultivate the cells for 4 days in BM-Cyclin 2
Repeat this cycle twice.