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Reagents and Media components

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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

Horse Serum: 500ml, Gibco-BRL # 16050-023

Geneticin (G418): Purchase from Gibco-BRL

Hygromycin: Purchase from Boehringer Mannheim

VacuCap Filters 0.2um sterile filter, 10/pkg. Gelman Sciences cat. # 4622

Tissue Culture dishes: usually Corning brand, available from HSC stores.

25X PBS

5g KCl

5g KH2PO4

29g Na2HPO4 2H2O

200g NaCl

make up to 1l with MQ water and autoclave.