Recipies for Cell Fusion
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<center> <b><font>Recipies for Cell Fusion</font> </b></center>
Phosphate-buffered saline (PBS) (0.01 M Na2HPO4 , 0.15 M NaCl, pH 7.4)
| sodium phosphate dibasic (Na2HPO4) | 0.87 g |
| sodium phosphate monobasic (NaH2PO4 x H2O) | 0.54 g |
| sodium chloride | 8.5 g |
| HPLC water | 1 liter |
Adjust 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 ml |
| NCTC-135 medium | 10% | 50 ml |
| Fetal bovine serum | 20 % | 100 ml |
| Antibiotic/antimycotic | 1% | 5 ml |
| Fungizone | 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 ml |
| Distilled water | 130 ml |
Disinfecting solution (for cleaning)
| Roccall II | 5 ml |
| HPLC water | 1 liter |
Disinfecting solution (for filling water pan)
| Roccall II | 7.5 ml |
| HPLC water | 4 liter |
HAT media
| HAT supplement, 500 x | 0.1 ml |
| DMEM Complete media | 50 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 x | 0.1 ml |
| DMEM Complete media | 50 ml |
(NOTE: some formulations are 50 x and should be used at the rate of 1 ml / 50 ml media)
