Recipe for ECPM1 media? - (Mar/16/2008 )

Hey ya'll,

I am trying some expression tests in e. coli and want to try some different media. I used to use ECPM1 all the time in my last lab, but didn't think to save the recipe. I remember it being a great media for those toughies. If anyone has the recipe for the media and the trace elements, I'd be susper-stoked.

~B

see this website: http://mrw.interscience.wiley.com/emrw/978...03/current/html

ECPM1 medium
4 g K2HPO4
1 g KH2PO4
1 g NH4Cl
2.4 g K2SO4
132 mg CaCl2·2H2O
10 ml trace element solution 1 (see recipe)
20 g Casamino Acids (enzymatic hydrolysate; Difco)
3 g yeast extract (Difco)
40 g glycerol
H2O to 800 ml

Mix ingredients, pour into fermentor, and sterilize by heat (see Basic Protocol 1, steps 5 to 8) adding balance of water to 1 liter in step 6. Add 2 ml of 1 M MgCl2·6H2O (sterilized by filtration through a membrane with 0.22-µm pores) as directed in step 12 of Basic Protocol 1. Once sterilized, the medium can be stored up to 1 week at 4°C.

Trace element solution 1
5 g EDTA
0.5 g FeCl3·6H2O
0.05 g ZnO
0.01 g CuCl2·2H2O
0.01 g Co(NO3)2·6H2O
0.01 g (NH4)6Mo7O24·4H2O
Dissolve EDTA in 700 ml water. Dissolve each element separately by adding just enough 10 N HCl to bring it into solution. Add each element in solution one after the other to the EDTA solution. Add water to 1 liter. Adjust pH to 7.0 with concentrated NaOH. Autoclave. Store in the dark at 4°C.