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Recipies for Cell Fusion |
| 点击： 作者： 来源： 时间： 2006-11-07 本站论坛 |
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Recipies for Cell Fusion
Phosphate-buffered saline (PBS) (0.01 M Na2HPO4 , 0.15 M NaCl, pH 7.4)
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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)
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