【成份】
1064
Solution A:
Trypticase peptone (BD BBL) 1.0 g
Mineral solution 50.0 ml
Trace element solution SL-10 1.0 ml
Rumen fluid
clarified 50.0 ml
Butyric acid 1.7 g
Na2SO4 2.8 g
Na-resazurin solution (0.1% w
) 0.5 ml
蒸馏水 830.0 ml
Solution B:
NaHCO3 3.5 g
蒸馏水 50.0 ml
Solution C:
Vitamin solution 1.0 ml
Solution D:
L-Cysteine-HCl·H2O 0.3 g
蒸馏水 10.0 ml
Solution E:
Na2S·9H2O 0.3 g
蒸馏水 10.0 ml
Add and dissolve ingredients of solution A
Mineral solution:
KH2PO4 10.0 g
MgCl2·6 H2O 6.6 g
NaCl 8.0 g
NH4Cl 8.0 g
CaCl2·2 H2O 1.0 g
蒸馏水 1.0 L
Trace element solution SL-10:
HCl (25%; 7.7 M) 10.0 ml
FeCl2·4 H2O 1.5 g
ZnCl2 70.0 mg
MnCl2·4 H2O 100.0 mg
H3BO3 6.0 mg
CoCl2·6 H2O 190.0 mg
CuCl2 ·2 H2O 2.0 mg
NiCl2·6H2O 24.0 mg
Na2MoO4·2H2O 36.0 mg
蒸馏水 990.0 ml
First dissolve FeCl2 in the HCl
then dilute in water
add and dissolve the other salts. Finally make up to 1000.0 ml.
Clarified rumen fluid:
Rumen fluid from cow or sheep is filtered through muslin
autoclaved at 121°C for 15 min and then centrifuged at 27
000g for 20 min. The supernatant is made anoxic by sparging with 100% N2 gas for 15 min
dispensed under same gas atmosphere into anoxic serum vials to 30% of volume and then stored frozen at -20°C.
Seven vitamins solution:
Vitamin B12 100.0 mg
p-Aminobenzoic acid 80.0 mg
D(+)-Biotin 20.0 mg
Nicotinic acid 200.0 mg
Calcium pantothenate 100.0 mg
Pyridoxine hydrochloride 300.0 mg
Thiamine-HCl x· 2 H2O 200.0 mg
蒸馏水 1.0 L