Cell Culture Solutions BB 9/15/05
Updated 6/15/07 JEB
Borate Buffer (4º)*
· 4.76 g Boric Acid (Cat # B-1934)
· 2.54 g Borax (Cat # B-0127)
· 1000 mL of ddH2O
· pH to 8.4 with NaOH
· Sterile Filter
Plating Media (4º) □
· 40 mL F12 (Cat # N8641)
· 40 mL FBS (Cat # sh30070.03)
· 420 mL Dulbecco Modified Eagle Medium (DMEM) W/O L-Glutamine (Cat # 11960)
· 1 mL of PS (Cat # 17-602E)
· 200 uL of 200 mM L-Glutamine (Cat # G7513)
D2C (4º) □
· 10 mL FBS (Cat # sh30070.03)
· 1 mL PS (Cat # 17-602E)
· 12.5 mL of 1M Hepes (Cat # H0887)
· 471.5 mL Dulbecco Modified Eagle Medium (DMEM) W/O L-Glutamine (Cat # 11960)
· 6.25 mL of 200 mM L-Glutamine (Cat # G7513)
Neurobasal (4º) □
· 1 bottle (500 mL) Neurobasal Media (Cat # 21103)
· 1 bottle (10 mL) 50X B27(Cat # 17504-044)
· P/S (Cat # 17-602E)
Stock Ara-C (500X) (-20º)*
· 100 mg Ara-C (Cat # C-6645)
· 6 mL sterile H2O
· Sterile Filter
· Aliquot 0.5 mL
Working Ara-C (4º)*
· 20µL Stock
· 10 mL sterile T.C. water
10% Formaldehyde (RT or 4°)*
· 5 mL of 37% Stock Solution
· 45 mL of PBS
Glucose Free Balanced Salt Solution*
· 5.26 g NaCl (Cat # S9625-5KG)
· 0.125 g KCl (Cat # P-9541)
· 66.6 mg CaCl2 (Cat # C-4901)
· 1.43 g Hepes (Cat # H-4034)
· 600 mL ddH2O
· Sterile Filter
· pH 7.3
PLO Δ
· 1 mg PLO : 2 mL Borate Buffer (Cat # P3655)
· Sterile filter
· Make up fresh each time it is used
HBSS (4º)*
· 8.4 g NaCl (Cat # S9625-5KG)
· 224 mg KCl (Cat # P-9541)
· 2.38 g Hepes (Cat # H-4034)
· 1.0 g Glucose (Cat # G-5250)
· 1000 mL Milli Q
· pH to 7.3 with NaOH
· Sterile Filter
Roccal II (10%) (RT)*
· 10% Benzalkonium Chloride (Cat # 234427 Sigma)
· 1.25% Ethyl Alcohol
· Dissolve in alcohol with stirring
· Add 88.75% Milli Q water
MEM/BSA/Hepes □
· 487.5 mL of clear MEM (Cat # 51200)
· 12.5 mL of 1 M Hepes (Cat # H0887)
· 50 mg of BSA (Cat # A7888)
· Sterile Filter
*=good for months, □=good for ~1 week, Δ= make fresh
Note: We are no longer heat inactivating our Fetal Bovine Serum