SLSM

SLSM is supplemented liquid sporulation media.  We use it after GNA plates in sporulation.

SLSM, to make 200mls:

20mls of 10% Potassium Acetate

2mls of 0.5% Zinc Acetate

2mls of 100X URA Supplement solution

1ml of 200X His Supplement solution

1ml of 200X Leu supplement solution

174ml ddH2O

Autoclave

Ingredient	Amount (ml)
10% K Acetate	20
0.5% Zn Acetate	2
100X URA Supp Soln	2
200X HIS Supp Soln	1
200X LEU Supp Soln	1
ddH2O	174

Stocks used:

10% Potassium Acetate: 20g K Acetate (bioWORLD 41600116-3) in 200ml total H2O, autoclaved

0.5% Zn Acetate:  1.196g Zn Acetate Dihydrate (SIGMA 383058-500G) in 200ml total H2O, autoclaved

100X URA Supp Soln: 200mg URA (bioWORLD 42190000-1) in 100ml total H2O, autoclaved

200X HIS Supp Soln: 0.4g HIS (bioWORLD 40800052-1) in 100ml total H2O, autoclaved

200X LEU Supp Soln: 0.6g LEU (SIGMA L8000-250G) in 100ml total H2O, autoclaved

OR

Alternate Protocol:

Add all of the ingredients in a clean, autoclaved bottle.

Add enough ddH2O to dissolve the ingredients and to make a solution of your desired volume (e.g. 200mL, 1L).

Dissolve completely, label the bottle (SLSM, YOUR INITIALS, DATE), then autoclave (ASK SOMEONE HOW TO DO THIS).

Ingredients	For a 200mL Solution	For a 1L Solution
Potassium Acetate	2 g	10 g
Zinc Acetate Dihydrate	0.0120 g	0.0598 g
L-URA	0.004 g	0.02 g
L-HIS	0.004 g	0.02 g
L-LEU	0.006 g	0.03 g