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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 -