alpha-Minimal Essential Medium with L-glutamine, without ribo-/deoxyribonucleosides.
Step1. 1 bottle of AlphaMEM 10L powder 8L dH2O mixture
Step2. NHCO3 22g dH2O 700ml mixture
Mix step1 and step2 together before filter. Go through 0.1 m filter (Gelman) make 450 ml of each bottle.
Medium (alpha-MEM) without bicarbonate. We make this ourselves from premixed powder and filter sterilize. Gibco-BRL cat # 12000-063
F11 Medium: available ready made from the Cancer Research Group (CRG).
Joklik's MEM: available ready made from the CRG.
Penicillin - Streptomycin: available from the CRG as (100 X Pen-Strep). It can also be purchased Gibco-BRL cat # 15070-014.
L-glutamine (100X): available from the CRG. or Gibco-BRL # 25030-016.
Fungizone (40X): Gibco-BRL cat # 15295-017.
Trypsin.EDTA (1X): Gibco-BRL cat # 25300-021.
Trypsin.EDTA (10X): available from the CRG or Gibco-BRL # 15400-013, or diluted 10x trypsin (GIBCO) with 1x PBS.
Calf Serum: 500ml Gibco-BRL # 16170-011.
Newborn Calf Serum: 500ml, Gibco-BRL # 16010-043
Fetal Bovine Serum: 500ml, Gibco-BRL # 16000-028 - Heat inactive FBS Aliquot 50ml FBS to falcon tube, put into the 55 C water bath for 1 hour, store in -20ºC.
- Charcoal filtered FBS
150ml FBS
12g Charcoal (sigma)
Mixed for 1 and half hour Go through 0.45 micron then 0.2 micron Filter