# 1200: ANAEROMYXOBACTER-MEDIUM


## Main sol. 1200

10	ml	**Solution A**	
1	ml	**Solution B**	
0.2	ml	**Solution C**	
0.2	ml	**Solution D ("red mix")**	
0.05	ml	**Solution E (electron acceptor)**	
0.2	ml	**Solution F: Vitamin solution according to Wolin and coauthors**	

1. Prepare solutions A, B, and D separately and anaerobically under 80% N2 + 20% CO2 and autoclave. Filter-sterilize solutions C, E and F and outgas with N2. After cooling, add about 1 ml solution B per 10 ml solution A, by this way adjusting pH to 7.2. Add 0.2 ml solution C, 0.2 ml solution D, 0.2 ml solution F and 0.05 ml solution E, each per 10 ml solution A.


## Solution A

1	g	NaCl	
0.5	g	MgCl2 x 6 H2O	
0.2	g	KH2PO4	
0.3	g	NH4Cl	
0.3	g	KCl	
15	mg	CaCl2 x 2 H2O	
0.4	g	Na-acetate	
1	ml	**Trace element solution SL-10 B**	
2	ml	**Selenite-tungstate solution**	
1	ml	Resazurin (1 g per L)	
900	ml	Distilled water	


## Solution B

2.5	g	NaHCO3	
50	ml	Distilled water	


## Solution C

385	mg	DL-Dithiothreitol	
50	ml	Distilled water	


## Solution D ("red mix")

37.5	mg	L-Cysteine	
40	mg	Na2S x H2O (dissolved separately in water)	
50	ml	Distilled water	


## Solution E (electron acceptor)

4	g	Disodium fumarate	
50	ml	Distilled water	


## Solution F: Vitamin solution according to Wolin and coauthors

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 element solution SL-10 B

1000	ml	Distilled water	
7.7	ml	HCl (25%)	
1.5	g	FeSO4 x 7 H2O	
70	mg	ZnCl2	
100	mg	MnCl2 x 4 H2O	
300	mg	H3BO3	
190	mg	CoCl2 x 6 H2O	
2	mg	CuCl2 x 2 H2O	
24	mg	NiCl2 x 6 H2O	
36	mg	Na2MoO4 x 2 H2O	


## Selenite-tungstate solution

0.5	g	NaOH	
3	mg	Na2SeO3 x 5 H2O	
4	mg	Na2WO4 x 2 H2O	
1000	ml	Distilled water