KCTC Media No. 1834 : SB48 cultivation medium

조성	용량	단위
Solution 1	5.0	ml
조성 용량 단위 NH_4Cl 33 g/L KCl 33 g/L MgCl_2·6H_2O 33 g/L CaCl_2·6H_2O 33 g/L
Solution 2	5.0	ml
조성 용량 단위 KH2PO4 33 g/L
Trace elements solution (mmol/l)	0.5	ml
조성 용량 단위 (NH_4)_2Fe(SO_4)_2·6H_2O 2.0 Na2SO4 2.0 CoCl_2·6H_20 1.0 NiCl_2·6H_2O 1.0 MnCl_2·4H_2O 0.5 ZnSO_4·7H_2O 0.5 Na2SeO3 0.5 Na2MoO4·2H_2O 0.1 Na2WO4·2H_2O 0.1 H3BO3 0.1 CuCl_2·2H_2O 0.01
Vitamin Solution (See KCTC medium No. 1335)	0.5	ml
NaCl	9	g
MgCl2·6H2O	2.0	g
Distilled water	500	ml
NaHCO3	1.0	g
10% NaOH solution	3.0	ml
pyruvate	10.0	mM
nitrate	10.0	mM

** ph : -

** memo : -

** 추가조치 사항 : The medium has to be prepared anaerobically, however no reducing agent or resazurin are added.
- Add solutions 1 (5 ml), 2 (5 ml), trace elements solution (0.5 ml), 9 g NaCl and 2 g MgCl2*6H2O to 500 ml of distilled water.
- Boil for 5 min and cool for 10 min under CO2 (100%) ? do not bubble
- add solution of vitamins ? 0.5 ml
- add and dissolve NaHCO3 ? 1 g (2 g/l)
- add 3 ml of 10% NaOH solution
- bubble CO2 (100 %) through the medium for ca. 2 min.
- Dispense by 10 ml to 17-ml Hungate tubes under CO2 (100%) flux.
- Autoclave at 1 atm for 1 hour.

Add pyruvate (10 mM) and nitrate (10 mM) from anaerobically prepared sterile stock solutions before inoculation. Incubate at 30oC for about 5 days.