KCTC Media No. 1412 : Modified Methanobacterium medium

조성	용량	단위
KH2PO4	0.5	g
MgSO47H2O	0.4	g
NaCl	0.4	g
NH4Cl	0.4	g
CaCl22H2O	0.05	g
Trace element solution SL-10(see medium no. 1136)	1.0	ml
Brain Heart Infusion (BD Difco)	6	g
Proteose peptone (BD Difco)	6	g
Yeast extract(OXOID)	2.0	g
Na-acetate	1.0	g
Na-formate	2.0	g
Sodium Resazurin (0.1% w/v)	0.5	ml
NaHCO3	4	g
Vitamins solution	1.0	ml
조성 용량 단위 Vitamin B12 100 mg p-Aminobenzoic acid 80 mg D(+)-Biotin 20.0 mg Nicotinic acid 200 mg Calcium pantothenate 100 mg Pyridoxine hydrochloride 300 mg Thiamine-HCl x 2 H2O 200 mg Distilled water 1.0 L
L-Cystein-HCl·H2O	0.5	g
Na2S·9H2O	0.5	g
Distilled water	1.0	L

** ph : 6.8-7.0

** memo : -

** 추가조치 사항 : 1. Dissolve ingredients except bicarbonate, vitamins, cysteine, and sulfide, then sparge
medium with 80% H2 and 20% CO2 gas mixture for 30 - 45 min to make it anoxic. Add and
dissolve bicarbonate and adjust pH to 6.5. Then dispense medium under 80% H2 and 20%
CO2 gas atmosphere into anoxic Hungate-type tubes or serum vials to 30% of their volume
and autoclave. Add vitamins, cysteine, and sulfide from sterile anoxic stock solutions
prepared under 100% N2 gas. The vitamin solution should be sterilized by filtration. Before
use, check the complete medium's pH and adjust it to 6.8 - 7.0, if necessary.
2. After growth has started and the culture is becoming turbid add sterile 80% H2 and 20%
CO2 gas mixture to 0.5 - 1 bar overpressure.

For KCTC4028, KCTC4029, KCTC25506 :
Supplement medium after autoclaving with 0.50 g/l coenzyme M (2-mercaptoethanesulfonic acid) and 0.30 g/l DTT (DL-Dithiothreitol) added from filter-sterilized anoxic stock solutions prepared under N2.
Omit sulfide and cysteine from the medium.