山(shan)東拓普(pu)生物工程有限公司

Shandong Tuopu Biol-Engineering Co.,Ltd

Anaerobranca gottschalkii Medium
(DSMZ Medium 895)
Composition 1070mL:
NaCl............................................................................................10.0g
(NH 4 ) 2 SO 4 ....................................................................................1.0g
K 2 HPO 4 .........................................................................................0.5g
L -Cysteine.....................................................................................0.5g
NH 4 Cl ...........................................................................................0.4g
Yeast extract................................................................................0.25g
Tryptone......................................................................................0.25g
Na 2 S 2 O 3 ·5H 2 O..............................................................................0.1g
MgSO 4 ·7H 2 O................................................................................0.1g
CaCl 2 ·2H 2 O.................................................................................0.05g
FeSO 4 ·7H 2 O...............................................................................2.0mg
Resazurin ...................................................................................0.5mg
Na2CO3 solution........................................................................50.0mL
Soluble starch solution.............................................................20.0mL
Trace elements solution ...........................................................10.0mL
Vitamin solution, 10 fold conc...................................................1.0mL
pH 9.4 ± 0.2 at 25°C
Trace Elements Solution:
Composition per liter:
MgSO4 ·7H2O................................................................................3.0g
Nitrilotriacetic acid.......................................................................1.5g
NaCl..............................................................................................1.0g
MnSO4 ·2H2O................................................................................0.5g
CoSO4 ·7H2O...............................................................................0.18g
ZnSO4 ·7H2O...............................................................................0.18g
CaCl2 ·2H2O...................................................................................0.1g
FeSO4 ·7H2O..................................................................................0.1g
NiCl2 ·6H2O...............................................................................0.025g
KAl(SO4）2 ·12H2O.......................................................................0.02g
H3BO3 .........................................................................................0.01g
Na2MoO4 ·4H2O ..........................................................................0.01g
CuSO4 ·5H2O...............................................................................0.01g
Na2SeO3 ·5H2O...........................................................................0.3mg
Preparation of Trace Elements Solution: Add nitrilotriacetic
acid to 500.0mL of distilled/deionized water. Dissolve by adjusting pH
to 6.5 with KOH. Add remaining components. Add distilled/deionized
water to 1.0L. Mix thoroughly.
Vitamin Solution:
Composition per 100.0mL:
Pyridoxine-HCl........................................................................10.0mg
Thiamine-HCl·2H 2 O..................................................................5.0mg
Riboflavin ..................................................................................5.0mg
Nicotinic acid.............................................................................5.0mg
D -Ca-pantothenate......................................................................5.0mg
p-Aminobenzoic acid.................................................................5.0mg
Lipoic acid .................................................................................5.0mg
Biotin .........................................................................................2.0mg
Folic acid....................................................................................2.0mg
Vitamin B 12 ................................................................................0.1mg
Preparation of Vitamin Solution:  Add components to distilled/
deionized water and bring volume to 100.0mL. Mix thoroughly. Sparge
with 80% H 2 + 20% CO 2 . Filter sterilize.
Na 2 CO 3 Solution:
Composition per 100.0mL:
Na 2 CO 3 .........................................................................................5.0g
Preparation of Na 2 CO 3 Solution:  Add Na 2 CO 3 to distilled/de-
ionized water and bring volume to 100.0mL. Mix thoroughly. Sparge
with 100% N 2 . Autoclave for 15 min at 15 psi pressure–121°C.
Soluble Starch Solution:
Composition per 50.0mL:
Starch, soluble...............................................................................5.0g
Preparation of Soluble Starch Solution:  Add starch to distilled/
deionized water and bring volume to 50.0mL. Mix thoroughly. Sparge
with 100% N 2 . Autoclave for 15 min at 15 psi pressure–121°C.
Preparation of Medium: Add components, except starch solution,
Na 2 CO 3 solution, and  L -cysteine, to distilled/deionized water and bring
volume to 1.0L. Mix thoroughly. Gently heat and bring to boiling. Cool
to 25°C while sparging with 100% N 2 . Add 0.5g  L -cysteine. Mix thor-
oughly. Distribute to anaerobe tubes or bottles. Autoclave for 15 min at
15 psi pressure–121°C. Cool to 25°C. Aseptically and anaerobically
add,  per liter of medium, 20.0mL sterile starch solution, and 50.0mL
sterile Na 2 CO 3 solution. Final pH is 9.3–9.5.
Use:  For the cultivation of Anaerobranca gottschalkii.