Cell Culture

1. Dulbecco's modified eagle's medium (DMEM, Cambrex BioScience, Belgium), supplemented with 10% fetal bovine serum (FBS, Hyclone, Belgium).

2. Penicillin (used at 100 U/mL final concentration) and streptomycin (used at 100 yg/ mL final concentration), stored at -20°C (100X stock from Cambrex BioScience, Belgium).

3. 100% Dimethylsulfoxide (DMSO, Merck, Germany), used at 2% final concentration.

4. Neomycin, stock prepared as 100 mg/mL in phosphate-buffered saline (PBS; see Subheading 2.2., item 1), filter-sterilized, aliquoted, stored at -20°C, and used at 400 yg/mL final concentration.

5. Puromycin, 1000X stock aliquoted and stored as above and added to 1 yg/mL final concentration.

0 0

Post a comment