KCTC Media No. 976 : DB CHARACTERIZATION MEDIUM NO. 2

조성	용량	단위
MDS salt water	833	ml
조성 용량 단위 NaCl 240 g MgCl_2·6H_2O 30.0 g MgSO_4·7H_2O 35 g KCl 7 g 1M CaCl_2 solution 5.0 ml
FeCl2 solution	1.0	ml
조성 용량 단위 25% HCl (7.7 M) 10.0 ml Distilled water 990 ml FeCl_2·4H_2O 1.5 g
Trace element solution SL-6	1.0	ml
조성 용량 단위 ZnCl_2 70 mg MnCl_2·4H_2O 100 mg H3BO3 6 mg CoCl_2·6H_2O 190 mg CuCl_2·2H_2O 2.0 mg NiCl_2·6H_2O 24 mg Na2MoO4·2H_2O 36 mg Distilled water 1.0 L
Peptone (Oxoid)	0.25	g
Yeast extract (BD)	0.05	g
Add components and bring volume to 980 ml with distilled water. For preparation of solid medium, add 20 g/L agar (if necessary, use washed agar as described below). Autoclave, and after cooling, add the following solutions (filter-sterilized or ·autoclaved) to the medium:
1M NH_4Cl	5.0	ml
Potassium phosphate buffer·	2.0	ml
조성 용량 단위 1M K2HPO4 83.4 ml 1M KH2PO4 16.6 ml
Vitamin solution	3.0	ml
조성 용량 단위 Biotin 3.0 mg Folic acid 4 mg Pyridoxine·HCl 50 mg Thiamine ·HCl 33 mg Riboflavin 10.0 mg Nicotinic acid 33 mg DL-Calcium pantothenate 17 mg Vitamin B12 17 mg Lipoic acid 10.0 mg p-Aminobenzoic acid 13 mg Distilled water 1.0 L
1 M Sodium pyruvate solution	10.0	ml
Readjust pH to 7.0 - 7.5 with a sterilized 10% Na2CO3 solution, if necessary. Washed agar: Stir agar in distilled water and stand still for 30 min. Then, remove supernatant by decantation. Repeat the washing at least twice.
Add components to distilled water and bring volume to 1.0 L. Adjust pH to 7.5 with 1 M Tris base.
Combine the two solutions and check that pH is close to 7.5. Add an equal volume of distilled water.

** ph : -

** memo : JCM 574 medium Add components and bring volume to 980 ml with distilled water. Autoclave, and after cooling, add the following solutions (filter-sterilized or *autoclaved) to the medium:
1 M NH_4_Cl 5.0 ml
Potassium phosphate buffer* 2.0 ml
Vitamin solution 3.0 ml
1 M Sodium pyruvate solution 10.0 ml

MDS salt water:
NaCl 240.0 g
MgCl2·6H2O 30.0 g
MgSO4·7H2O 35.0 g
KCl 7.0 g
1 M CaCl2 solution 5.0 ml
Add components to distilled water and bring volume to 1.0 L. Adjust pH to 7.5 with 1 M Tris base.

Potassium phosphate buffer:
1 M K2HPO4 83.4 ml
1 M KH2PO4 16.6 ml
Combine the two solutions and check that pH is close to 7.5. Add an equal volume of distilled water.

Vitamin solution:
Biotin 3.0 mg
Folic acid 4.0 mg
Pyridoxine·HCl 50.0 mg
Thiamine·HCl 33.0 mg
Riboflavin 10.0 mg
Nicotinic acid 33.0 mg
DL-Calcium pantothenate 17.0 mg
Vitamin B12 17.0 mg
p-Aminobenzoic acid 13.0 mg
Lipoic acid 10.0 mg
Distilled water 1.0 L

FeCl2 solution:
25% HCl (7.7 M) 10.0 ml
FeCl2·4H2O 1.5 g
Distilled water 990.0 ml

Trace element solution:
ZnCl2 70.0 mg
MnCl2·4H2O 100.0 mg
H3BO3 6.0 mg
CoCl2·6H2O 190.0 mg
CuCl2·2H2O 2.0 mg
NiCl2·6H2O 24.0 mg
Na2MoO4·2H2O 36.0 mg
Distilled water 1.0

** 추가조치 사항 : -