#9 MAKING SCC CELL MEDIA:

Powder: Dulbecco's modified eagle's media: Nutrient mix F12 Ham's = DMEM/F12 1:1
Hyclone (SH30004)




To make 500ml DMEM:F12 1:1 Media:


Make with stirring in lab mixing plate (will sterilize later)

1.      400ml water room temp

2.      7.8 g DMEM: F12 powder (4C)

3.      stir these until powder dissolves first

4.      Then add 8ml 7.5% sodium bicarbonate (NaHCO3)—tissue culture
grade/sterile, stir, will change pH and color = will be pH ~6.8 = ok
(allowance for pH between 6.9-7.3)

5.      Add water to 500ml total

6.      Sterile filter using vaccum and a 0.2um filter bottle attachment

7.      Add 10% FBS (50ml), 1% PS (5.5ml), 1% fungicide (5.5ml) in Tissue
Culture Hood

8.      Add also in Tissue Culture Hood: hydrocortisone 400ng/mL. From 4
ug/ul = 200ul = 200ug.




Keep sterile and happy culturing!