# 986: PARAMALEDIVIBACTER MEDIUM


## Main sol. 986

30	g	Sea Salt (SIGMA)	
1	g	NH4Cl	
0.3	g	K2HPO4	
0.3	g	KH2PO4	
0.5	g	Peptone	
0.5	g	Yeast extract	
0.5	ml	Sodium resazurin (0.1% w/v)	
2	g	NaHCO3	
4	g	D-Glucose	
1	ml	**Wolin's vitamin solution (10x)**	
0.25	g	L-Cysteine HCl x H2O	
0.25	g	Na2S x 9 H2O	
1000	ml	Distilled water	

1. Dissolve ingredients (except bicarbonate, glucose, vitamins, cysteine and sulfide), sparge medium with 100% N2 gas for 30 - 45 min to make it anoxic. Dispense medium under same gas atmosphere into anoxic Hungate-type tubes or serum vials and autoclave. Add vitamins, glucose, sulfide and cysteine from sterile anoxic stock solutions prepared under 100% N2 gas and bicarbonate from a sterile anoxic stock solution prepared under 80% N2 and 20% CO2 gas mixture. Vitamins should be sterilized by filtration. The pH of the complete medium should be 7.0.


## Wolin's vitamin solution (10x)

20	mg	Biotin	
20	mg	Folic acid	
100	mg	Pyridoxine hydrochloride	
50	mg	Thiamine HCl	
50	mg	Riboflavin	
50	mg	Nicotinic acid	
50	mg	Calcium D-(+)-pantothenate	
1	mg	Vitamin B12	
50	mg	p-Aminobenzoic acid	
50	mg	(DL)-alpha-Lipoic acid	
1000	ml	Distilled water