allow solution to cool to room temperature before final ph adjustment dilute to a final volume of 1l te buffer 10 0 mm tris buffer 1 0 m m edta ph 7 4 lb luria bertani 10 g bacto tryptone 5 g bacto yeast extract 8 g nacl in 1 l of water adjusted to ph 7 5 with naoh______________________________________ypd medium 1 bacto yeast extract 2 bacto peptone 2 dextroseim sub 1 minimal medium im salts at a final concentration of 36 7 mm kh sub 2 po sub 4 22 7 mm nh sub 4 sub 2 so sub 4 2 0 mm mgso sub 4 u00b7 7h sub 2 o 6 7 mm kcl 0 7 mm cacl sub 2 u00b7 2h sub 2 o prepared as a 10x stock solution and autoclaved trace salts at a final concentration of 0 2 u03bcm cuso sub 4 u00b7 5h sub 2 o 1 25 u03bcm ki 4 5 u03bcm mnso sub 4 u221a h sub 2 o 2 0 u03bcm namoo sub 4 u00b7 2h sub 2 o 0 75 u03bcm h sub 3 bo sub 3 17 5 u03bcm znso sub 4 u00b7 7h sub 2 o 44 5 u03bcm fecl sub 3 u00b7 6h sub 2 o prepared as a 400x stock solution and filter sterilized 0 4 u03bcgl biotin and 2 dextrose ______________________________________ 5 g bacto dextrose 1 g sodium chloride 3 5 g dipotassium phosphate 3 68 g monopotassium phosphate 1 32 g pichia pastoris was grown in ypd rich or im 1 minimal media e coli was cultured in either lb medium or 2b medium 0 2 nh 4 po 4 1.