# 1148: TINDALLIA MEDIUM


## Main sol. 1148

30	g	NaCl	
0.2	g	KCl	
0.2	g	K2HPO4	
0.1	g	MgCl2 x 6 H2O	
1	g	NH4Cl	
0.5	g	Yeast extract	
5	g	Peptone	
1	ml	**Trace element solution (mg per 200 ml)**	
1	mg	Resazurin	
1000	ml	Distilled water	

1. Prepare the medium omitting Na2CO3, NaHCO3, Na2S x 9 H2O, and Vitamin solution.
2. Boil the medium and cool under nitrogen, add Na2CO3 and NaHCO3.

24	g	NaHCO3	
2.76	g	Na2CO3	

3. Adjust pH to 9.5-10.0 with 10M KOH.
4. Dispense into tubes or bottle fitted with rubber stoppers.
5. Autoclave at 121 °C for 15 min.
6. Add Na2S x 9 H2O (neutralized) and Vitamin solution from sterile stock solutions

0.4	g	Na2S x 9 H2O (from neutralized stock solution)	
2	ml	**Wolin's vitamin solution**	


## Trace element solution (mg per 200 ml)

720	mg	MnCl2 x 4 H2O	
400	mg	Fe(NH4)2(SO4)2 x 12 H2O	
200	mg	FeSO4 x 7 H2O	
200	mg	CoCl2 x 6 H2O	
200	mg	ZnSO4 x 7 H2O	
20	mg	Na2MoO4 x 2 H2O	
100	mg	NiCl2	
20	mg	CuSO4 x 5 H2O	
20	mg	AlK(SO4)2 x 12 H2O	
20	mg	H3BO3	
5	ml	HCl (concentrated)	
200	ml	Distilled water	


## Wolin's vitamin solution

2	mg	Biotin	
2	mg	Folic acid	
10	mg	Pyridoxine hydrochloride	
5	mg	Thiamine HCl	
5	mg	Riboflavin	
5	mg	Nicotinic acid	
5	mg	Calcium D-(+)-pantothenate	
0.1	mg	Vitamin B12	
5	mg	p-Aminobenzoic acid	
5	mg	(DL)-alpha-Lipoic acid	
1000	ml	Distilled water