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Culture Medium (Current)
Composition of commercially prepared WIF-B Medium (updated 3-15-05)
Reagent |
Source
& Cat No |
Quantity |
F-12
Coon’s modification |
Biosource P114-500 |
500
ml |
Serum |
Gemini:
screened |
25
ml |
Glutamax |
GIBCO 35030-061 |
5
ml |
10,000
u Penicillin 10
mg Streptomycin (100X)** |
SIGMA P3539 |
5ml |
Amphotercin
(Fungizone) |
SIGMA A2942 |
1
ml |
HAT
(1000X) |
Traditional
Cassio receipe |
0.5 |
** This is a powder, may want to consider the liquid form as a precaution for endotoxin toxicity
Summary of F-12 Medium components
Component (mg/L) | ||||
Inorganic salts | Cassio (mg/L) | Biosource P114-500 | Irvine Scientific/ 9046 | SIGMA F6636 |
CaCl2 2H20 | 135.2 | 125 | 125 | 165 |
CuSO4 5H20 | 0.002 | 0.002 | 0.002 | 0.0025 |
FeSO4 7H20 | 0.8 | 0.8 | 0.8 | 0.834 |
KCl | 284 | 305 | 305 | 305 |
KH2PO4 | 59.4 | 68 | 68 | 61.24 |
MgCl2 6H20 | 105.5 | 106 | 50 | 46.659 |
MgSO4 7H20 | 39.3 | 104 | 51 | 25.28 |
NaCl | 7550 | 7530 | 7530 | 7517 |
NaHCO3 | 2744 | 2500 | 2500 | - ( added 2700) |
Na2HPO4 7 H20 | 168.4 | 250 | 132.4 | 132.4 |
ZnSO4 7 H20 | 0.144 | 0.14 | 0.14 | 0.863 |
Other | ||||
Ascorbic Acid | 15 | 45 | 45 | 15 |
Glucose (dextrose) | 1802 | 2000 | 2000 | 1802 |
Hypoxanthine | 4 | 4 | 5.4 | 4.04 |
Linoleic acid | 0.08 | 0.09 | - | 0.09 |
Lipoic | 0.2 | 0.2 ( Thioctic acid) | 0.2 (Thioctic acid) | 0.2 (Thioctic acid) |
Phenol Red | 1.2 | 1.25 | 1.25 | 1.25 |
Putrescine 2HCl | 0.3 | 0.3 | 0.3 | 0.161 |
Sodium Pyruvate | 220 | 220 | 220 | 220 |
Thymidine | 0.7 | 0.7 | 0.7 | 0.7 |
Amino Acids | ||||
L-Alanine | 18 | 18 | 18 | 18 |
L Arginine-HCl | 422 | 422 | 422 | 422 |
L Asparagine | 26 | 30 | 30 | 30 |
L Aspartic Acid | 26 | 26 | 26 | 26 |
L Cysteine HCl | 64 | - | - | 70.26 |
L Glutamic Acid | 30 | 30 | 30 | 30 |
L Glutamine | 292 | - | - | 292 |
Glycine | 16 | 16 | 16 | 16 |
L Histidine HCl | 38 | 42 | 42 | 42 |
L Isoleucine | 8 | 7.8 | 7.8 | 7.8 |
L Leucine | 26 | 26.2 | 26 | 26.2 |
L Lysine HCl | 74 | 73 | 73 | 73 |
L Methionine | 8 | 9 | 9 | 9 |
L Phenylalanine | 10 | 10 | 10 | 10 |
L Proline | 70 | 70 | 70 | 70 |
L Serine | 22 | 21 | 21 | 21 |
L Threonine | 24 | 23.8 | 24 | 23.8 |
L Tryptophane | 4 | 4 | 4 | 4 |
L Tyrosine | 10 | 11 | 16 | 15.86 |
L Valine | 24 | 23.4 | 23 | 23.4 |
Vitamins | ||||
Biotin | 0.07 | 0.07 | 0.007 | 0.0073 |
D-Ca pantothenate | 0.5 | 0.5 | 0.5 | 0.238 (hemi calcium) |
Choline choloride | 14 | 13.8 | 14 | 13.96 |
Folic Acid | 1 | 1 | 1 | 1.32 |
i-inositol | 18 | 36 | 36 | 18.02 (Myo-inosotol) |
Niacinamide | 0.04 | 0.04 | 0.04 | 0.04 |
Pyridoxine HCl | 0.06 | 0.06 | 0.06 | 0.06 |
Riboflavin | 0.04 | 0.04 | 0.04 | 0.04 |
Thiamine HCl | 0.3 | 0.2 | 0.28 | 0.337 |
Vitamin B 12 | 1 | 1 (cyanocobalamin) | 1 | 1.8 |
L cystine HCl | - | 48 | 63 | - |