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山東拓普生物工程有限公司

Shandong Tuopu Biol-Engineering Co.,Ltd

Anaerocellum Medium
Composition per liter:
Cellobiose or starch ......................................................................5.0g
NaHCO3 ........................................................................................1.5g
Na2S·9H2O....................................................................................0.5g
Yeast extract..................................................................................0.5g
CaCl2 ·2H2O.................................................................................0.33g
KCl..............................................................................................0.33g
KH2PO4 .......................................................................................0.33g
MgCl2 ·6H2O................................................................................0.33g
NH4Cl .........................................................................................0.33g
Resazurin ...................................................................................0.5mg
NaHCO3 solution...................................................................100.0mL
Cellobiose or starch solution....................................................50.0mL
Vitamin solution.......................................................................10.0mL
Na2S·9H2O solution.................................................................10.0mL
Trace elements solution SL-10 ..................................................1.0mL
pH 7.1–7.3 at 25°C
NaHCO3 Solution:
Composition per 100.0mL:
NaHCO3 ........................................................................................5.0g
Preparation of NaHCO 3 Solution:  Add NaHCO 3 to distilled/de-
ionized water and bring volume to 100.0mL. Mix thoroughly. Filter
sterilize. Gas under 80% N 2 + 20% CO 2 .
Cellobiose or Starch Solution:
Composition per 50.0mL:
Cellobiose or starch ......................................................................5.0g
Preparation of Cellobiose or Starch Solution:  Add cellobiose or
starch to distilled/deionized water and bring volume to 50.0mL. Mix
thoroughly. Filter sterilize. Gas under 100% N 2 .
Na 2 S·9H 2 O Solution:
Composition per 10.0mL:
Na 2 S·9H 2 O....................................................................................0.5g
Preparation of Na 2 S·9H 2 O Solution:  Add Na 2 S·9H 2 O to dis-
tilled/deionized water and bring volume to 10.0mL. Mix thoroughly.
Gas under 100% N 2 . Autoclave for 15 min at 15 psi pressure–121°C.
Vitamin Solution:
Composition per liter:
Pyridoxine·HCl........................................................................10.0mg
Calcium  DL -pantothenate...........................................................5.0mg
Lipoic acid .................................................................................5.0mg
Nicotinic acid.............................................................................5.0mg
p-Aminobenzoic acid.................................................................5.0mg
Riboflavin ..................................................................................5.0mg
Thiamine·HCl ............................................................................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 1.0L. Mix thoroughly. Gas under
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
Na2MoO 4 ·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: Add components, except NaHCO 3 solu-
tion, cellobiose or starch solution, and Na 2 S·9H 2 O solution, to distilled/
deionized water and bring volume to 830.0mL. Mix thoroughly. Gently
heat and bring to boiling. Continue boiling for 3–4 min. Allow to cool to
room temperature under 80% N 2 + 20% CO 2 . Distribute into bottles un-
der 80% N 2 + 20% CO 2 . Autoclave for 15 min at 15 psi pressure–121°C.
Aseptically and anaerobically add sterile NaHCO 3 solution, sterile cello-
biose or starch solution, and sterile Na 2 S·9H 2 O solution. Mix thorough-
ly.
Use: For the cultivation and maintenance of Anaerocellum thermophi-
lum and Dictyoglomus turgidus.