# 246a: REACTIVATION WITH LIQUID MEDIUM 246


## Main sol. 246a

1. Rehydrate and grow lyophilized cells from the ampoule in liquid medium 246. Subsequent subculturing may be carried out in liquid medium or on agar plates.

10	g	Beef extract	
10	g	Peptone	
20	g	Agar	
250	ml	Tap water	
750	ml	Sea water	

2. Dissolve beef extract and peptone by heating in tap water, adjust pH to 7.8 and boil for 10 min. Readjust pH to 7.3. Add agar and autoclave at 121°C for 20 min. Cool to 50°C and add warm (50°C) sterile sea water. Liquid medium without agar should be combined when cooled to room temperature.
3. Natural sea water is stored in the dark for at least three weeks to "age". If natural sea water is not available use artificial sea water.


## Artificial sea water

28.13	g	NaCl	
0.77	g	KCl	
1.6	g	CaCl2 x 2 H2O	
4.8	g	MgCl2 x 6 H2O	
0.11	g	NaHCO3	
3.5	g	MgSO4 x 7 H2O	
1000	ml	Distilled water