Phosphate-buffered saline (PBS) (0.01 M Na2HPO4 , 0.15 M NaCl, pH 7.4):sodium phosphate dibasic (Na2HPO4) 0.87 gsodium phosphate monobasic (NaH2PO4 x H2O) 0.54 gsodium chloride 8.5 gHPLC water 1 literAdjust the pH to 7.4 with 1 N NaOH or 1 M HCl.
Dubelcco Modified Eagle Medium (Complete Media):DMEM (1x liquid tissue culture medium with 4.5 g/l glucose) 70 % 500 mlNCTC-135 medium 10% 50 mlFetal bovine serum 20 % 100 mlAntibiotic/antimycotic 1% 5 mlFungizone 1 % 5 ml
1. Add 50 ml of NCTC-135 medium to a bottle of unopened DMEM media. Mix and transfer 50 ml to a bottle labelled “Washing Media.”
2. Add 100 ml of fetal bovine serum, 5 ml antibiotic and 5 ml fungizone to the 500 ml bottle of DMEM media. Mix, label the container “Complete Media”, and store in the refrigerator.
Washing Media:DMEM media, unopened 500 ml NCTC-135 media 50 ml(Note: Washing media is usually aquired by removing media before the addition of fetal calf serum. Fetal calf serum is expensive and necessary for cell growth.)
70% ethyl alcohol:95% ethyl alcohol 370 mlDistilled water 130 ml
Disinfecting solution (for cleaning):Roccall II 5 mlHPLC water 1 liter
Disinfecting solution (for filling water pan):Roccall II7.5 mlHPLC water4 literHAT mediaHAT supplement, 500 x0.1 mlDMEM Complete media50 ml(Note: some formulations are 50 x and should be used at the rate of 1 ml / 50 ml media)
HT media:HT supplement, 500 x0.1 mlDMEM Complete media50 ml(NOTE: some formulations are 50 x and should be used at the rate of 1 ml / 50 ml media)
