# 1361: PLANCTOMYCETALES BACTERIUM MEDIUM


## Main sol. 1361

0.14	g	KH2PO4	
0.2	g	MgCl2 x 6 H2O	
0.15	g	CaCl2 x 2 H2O	
0.54	g	NH4Cl	
1	ml	**Trace elements Solution**	
0.18	g	Glucose	
0.2	g	Yeast extract	
1	ml	**Vitamin solution**	
0.5	ml	Resazurin (0.1 v/v)	
2.5	g	NaHCO3	
0.3	g	Na2S x 9 H2O	
0.3	g	Cysteine HCl	
1000	ml	Distilled water	

1. Prepare the medium without yeast extract, glucose, NaHCO3, cysteine hydrochloride, vitamin solution and Na2S x 9 H2O. Boil and then cool under a gas atmosphere of N2:CO2 (80:20 v/v). Dispense the medium into Hungate tubes or serum bottles under gassing with N2:CO2 (80:20 v/v) and add the NaHCO3. The final pH should be 7.0. Autoclave the medium and add from sterile stock solutions yeast extract, glucose, cysteine hydrochloride, vitamin solution and Na2S x 9 H2O.The final pH should be 7.0.


## Vitamin solution

2	mg	Biotin	
2	mg	Folic acid	
10	mg	Pyridoxine hydrochloride	
5	mg	Thiamine-HCl x 2 H2O	
5	mg	Riboflavin	
5	mg	Nicotinic acid	
5	mg	D-Calcium pantothenate	
0.1	mg	Vitamin B12	
5	mg	p-Aminobenzoic acid	
5	mg	Lipoic acid	
1000	ml	Distilled water	


## Trace elements Solution

12.8	g	Nitrilotriacetic acid (NTA)	
1.35	g	FeCl3 x 6 H2O	
0.1	g	MnCl2 x 4 H2O	
0.024	g	CoCl2 x 6 H2O	
0.1	g	CaCl2 x 2 H2O	
0.1	g	ZnCl2	
0.025	g	CuCl2 x 2 H2O	
0.01	g	H3BO3	
0.024	g	Na2MoO4 x 2 H2O	
0.12	g	NiCl2 x 6 H2O	
4	mg	Na2SeO3 x 5 H2O	
1000	ml	Distilled water