KCTC Media No. 917 : MODIFIED BALCH'S METHANOGENS MEDIUM

조성	용량	단위
KH2PO4	0.3	g/L
K2HPO4	0.3	g/L
(NH4)2SO4	0.3	g/L
NaCl	0.6	g/L
MgSO2·7H2O	0.13	g/L
FeSO2·7H2O	0.002	g/L
CaCl2·2H2O	0.008	g/L
Yeast extract (Difco)	2.0	g/L
Trypticase peptone (BBL)	2.0	g/L
Sodium acetate	2.5	g/L
Sodium formate	2.5	g/L
Trace minerals (see Medium No. 918)	10	ml/L
조성 용량 단위 Nitrilotriacetic acid 1.5 g MgSO4·7H2O 3.0 g MnSO4·xH2O 0.5 g NaCl 1.0 g FeSO4·7H2O 0.1 g CoSO4·7H2O 0.1 g CaCl2·2H2O 0.1 g ZnSO4·7H2O 0.1 g CuSO4·5H2O 0.01 g AlK(SO4)2 0.01 g H3BO3 0.01 g Na2MoO4·2H2O 0.01 g Distilled water 1.0 L
Trace vitamins (see Medium No. 1335)	10	ml/L
조성 용량 단위 Biotin 2.0 mg Folic acid 2.0 mg Pyridoxine·HCl 10.0 mg Thiamine·HCl 5.0 mg Riboflavin 5.0 mg Nicotinic acid 5.0 mg Calcium pantothenate 5.0 mg Vitamin B12 0.1 mg p-Aminobenzoic acid 5.0 mg Lipoic acid 5.0 mg Distilled water 1.0 L
NaHCO3	5.0	g/L
Na2S·9H2O	0.5	g/L
L-Cysteine·HCl·H2O	0.5	g/L
Resazurin	0.001	g/L
Distilled water	1.0	L
Fatty acid mixture	20.0	ml/L
조성 용량 단위 Valeric acid 0.5 g Isovaleric acid 0.5 g α-Methylbutyric acid 0.5 g Isobutyric acid 0.5 g Distilled water 20.0 ml
coenzyme M solution (0.01% 2-mercaptoethanesulfonic acid, filter-sterilized)	1.0	ml/L

** ph : -

** memo : -

** 추가조치 사항 : Mix ingredients, except NaHCO3, L-Cysteine·HCl·H2O and Na2S·9H2O, and bring to a boil for 5 - 10 sec. Cool down under a stream of H2-CO2 (80:20), and add NaHCO3 to the medium. Dispense the medium into suitable culture vessels (e.g., 20 ml of the medium in 120 ml serum bottles) under a stream of H2-CO2 (80:20) and seal with butyl rubber stoppers. Separately autoclave cysteine·HCl and Na2S·9H2O as 5% solutions, respectively, under a N2 atmosphere. Prior to inoculation, aseptically and anaerobically add the cysteine·HCl and Na2S·9H2O solutions. Pressurize the inoculated vessels to 200 kPa H2-CO2 (80:20).


Adjust pH to 6.7-7.0 Prepare the medium anaerobically under a gas atmoshpere of 80 % H_2 _and 20 % CO_2_. Pressurize up to 2 atm.