KCTC Media No. 932 : MOPS Buffered Medium for Dehalococcoides sp.

조성	용량	단위
Basal Medium
조성 용량 단위 Salt solution (See Below) 20.0 ml Trace element solution (See Below) 1.0 ml Se/Wo solution (See Below) 1.0 ml Resazurin solution (See Below) 0.25 ml Sodium acetate 0.41 g D.W 956.5 ml
Boil and cool down to room temperature under N2, then add the following components:
Na2S·9H2O	0.2	mM
L-cysteine	0.2	mM
Autoclave medium at 121ºC and after the medium has cooled to room temperature add the following stock solutions:
NaHCO3 (See Below)	1.0	ml
MOPS (See Below)	20.0	ml
Ti(III)NTA(See Below)	20.0	ml
3-vitamin solution(See Below)	0.5	ml
6-vitamin solution (See Below)	1.0	ml
Vitamin B12 solution (See Below)	1.0	ml
Dispense anaerobically under 100% nitrogen into appropriate vessel.

** ph : -

** memo :

** 추가조치 사항 : **Salt solution
NaCl……………………………………………..50.0 g
MgCl2.6H2O…………………………………….20.5 g
NH4Cl……………………………………………13.5 g
KCl……………………………………………….26.0 g
CaCl2.2H2O……………………………………..0.75 g
KH2PO4………………………………………….10.0 g
DI Water…………………………………………1000 ml
Store at room temperature.

**Trace element solution
HCl (25% w/w)………………………………….10 ml
FeCl2.4H2O……………………………………...1.5 g
H3BO3……………………………………………6.0 mg
CoCl2.6H2O……………………………………..190.0 mg
MnCl2.4H2O……………………………………..100.0 mg
ZnCl2……………………………………………..70.0 mg
Na2MoO4. 2H2O…………………………………36.0 mg
NiCl2
.6H2O………………………………………..24.0 mg
CuCl2
.2H2O………………………………………2.0 mg
DI Water.…………………………………………1000 ml
Store at room temperature in the dark.

**Se/Wo solution
NaOH…………………………………………….0.5 g
Na2SeO3
.5H2O………………………………….6.0 mg
Na2WO4
.2H2O…………………………………..8.0 mg
DI Water…………………………………………1000 ml
Store at room temperature in the dark.

**Resazurin solution
Resazurin………………………………………..0.1 g
DI Water………………………………………….100 ml

**Stock solutions (add aseptically)
NaHCO3 1M autoclaved
MOPS, 1M pH adjusted to 7.45, filter sterilized
Ti(III)NTA, 25mM, filter sterilized

**3-vitamin solution
Folic acid…………………………………………1.0 mg
Riboflavin…………………………………………2.5 mg
DL-6,8-thioctic……………………………………2.5 mg
DI Water…………………………………………..250.0 ml
Filter sterilize: store at 4oC in the dark.

**6-vitamin solution
Biotin………………………………………………1.0 mg
4-aminobenzoic acid…………………………….5.0 mg
Pantothenate……………………………………..1.0 mg
Pyridoxamine……………………………………..25.0 mg
Nicotinic acid……………………………………..10.0 mg
Thiamine………………………………………….2.0 mg
DI Water………………………………………….100.0 ml
Filter sterilize: store at 4oC in the dark.

**Vitamin B12 solution
Vitamin B12…………………………………..….2.5 mg
DI Water………………………………………….100.0 ml
Filter sterilize: store at 4oC in the dark.