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山東(dong)拓普(pu)生物工程有限(xian)公司

Shandong Tuopu Biol-Engineering Co.,Ltd

Anaerofilum Medium
Composition per liter:
NaHCO3 ........................................................................................4.0g
Sodium formate.............................................................................2.0g
Sodium acetate..............................................................................1.0g
Yeast extract..................................................................................1.0g
L -Cysteine·HCl..............................................................................0.5g
KH2PO4 .........................................................................................0.5g
Na2S·9H2O....................................................................................0.5g
MgSO4 ·7H2O............................................................................... 0.4g
NaCl..............................................................................................0.4g
NH4Cl ...........................................................................................0.4g
CaCl2 ·2H2O.................................................................................0.05g
FeSO4 ·7H2O...............................................................................2.0mg
Resazurin...................................................................................1.0mg
Glucose solution......................................................................20.0mL
Fatty acid mixture....................................................................20.0mL
Trace elements solution SL-10..................................................1.0mL
pH 6.7 ± 0.2 at 25°C
Fatty Acid Mixture:
Composition per 20.0mL:
α-Methylbutyric acid....................................................................0.5g
Isobutyric acid ..............................................................................0.5g
Isovaleric acid...............................................................................0.5g
Valeric acid...................................................................................0.5g
Preparation of Fatty Acid Mixture : Add components to distilled/
deionized water and bring volume to 20.0mL. Mix thoroughly. Adjust
pH to 7.5 with concentrated NaOH.
Glucose Solution:
Composition per 20.0mL:
D -Glucose....................................................................................50.0g
Preparation of Glucose Solution:  Add glucose to distilled/deion-
ized water and bring volume to 20.0mL. Mix thoroughly. Sparge with
100% N 2 . Autoclave for 15 min at 15 psi pressure–121°C.
Trace Elements Solution SL-10:
Composition per liter:
FeCl2 ·4H2O...................................................................................1.5g
CoCl2 ·6H2O...........................................................................190.0mg
MnCl2 ·4H2O ..........................................................................100.0mg
ZnCl2 ........................................................................................70.0mg
Na2MoO4 ·2H2O.......................................................................36.0mg
NiCl2 ·6H2O..............................................................................24.0mg
H3BO3 ........................................................................................6.0mg
CuCl2 ·2H2O...............................................................................2.0mg
HCl (25% solution)..................................................................10.0mL
Preparation of Trace Elements Solution SL-10: Add FeCl 2 ·4H 2 O
to 10.0mL of HCl solution. Mix thoroughly. Add distilled/deionized
water and bring volume to 1.0L. Add remaining components. Mix thor-
oughly.
Preparation of Medium: Prepare and dispense medium anaerobi-
cally under 80% H 2 + 20% CO 2 . Add components, except glucose so-
lution, to distilled/deionized water and bring volume to 980.0mL. Mix
thoroughly. Adjust pH to 6.7. Sparge with 80% H 2 + 20% CO 2 . Auto-
clave for 15 min at 15 psi pressure–121°C. Aseptically and anaerobi-
cally add 20.0mL of sterile glucose solution. Aseptically and
anaerobically distribute into sterile tubes or bottles.
Use: For the cultivation of Anaerofilum agile and Anaerofilum pentos-
ovorans.