recipe for M63 minimal medium

Sergio sergioal at NOSPAMbbm1.ucm.es
Wed Jul 19 06:18:31 EST 2000


This medium lacks NaCl.

per liter:

KH2PO4            13.6 g  (100mM)
(NH4)2SO4        2 g       (15 mM)
FeSO4·7H2O     0,5 mg (1,7 uM)
Adjust pH to 7.0 with KOH

You can prepare a 10X or 5X stock, but i have experienced some precipitation
of the FeSO4 when using the 10X stock.

After autoclaving: add 1 ml of a 1 M solution of MgSO4·7H2O (directly in the
1X medium, not in the 10X).

Carbon source: 10 ml/l of a 20% v/v solution (final concentration: 0,2 %).
Sometimes I use even a 0,4% v/v carbon source, usually when i want a fast
and high density growing cells.

Vitamins should be added to a final concentration of 1ug/ml. (depends on the
strain, but i often add B1 and B12)

i hope this helps.

Sergio

sabine schill wrote:

> I want to try a secB null strain (Kumamoto, 1985) for expression, which
> is recommended to grow in M63 minimal medium. Unfortunately I don't have
> the reference "Miller, J.H., 1972. Experiments in molecular genetics.
> Cold Spring Harbor Laboratory, Cold Spring Harbor, N.Y.". Does anybody
> has a copy for me from this recipe or can give me another, which works
> too?
>
> Thanks a lot,
> Sabine
>
> ---






More information about the Methods mailing list