Neocallimastix Medium (ANO2 Fungus II)

[所属分类:培养基配方] [发布时间:2021-6-17] [发布人:网站管理员2] [阅读次数:] [返回]

Neocallimastix Medium
(ANO2 Fungus II)

山东拓普生物工程有限公司 培养基配方 http://www.topbiol.com

Composition per liter:

NaHCO 3 ........................................................................................6.6g
Cellobiose .....................................................................................2.0g
Glucose .........................................................................................2.0g
Maltose..........................................................................................2.0g
Starch ............................................................................................2.0g
PIPES (piperazine-N,N′-
bis[2-ethanesulfonic acid]) buffer ..........................................1.0g
Trypticase™ peptone.................................................................. 1.0g
Yeast extract..................................................................................1.0g
L -Cysteine·HCl·H 2 O .....................................................................0.5g
KH 2 PO 4 .........................................................................................0.5g
MgCl 2 ·6H 2 O..................................................................................0.4g
NaCl..............................................................................................0.4g
NH 4 Cl ...........................................................................................0.4g
Na 2 S·9H 2 O....................................................................................0.1g
CaCl 2 ·2H 2 O..............................................................................50.0mg
Resazurin ...................................................................................1.0mg
Rumen fluid ...........................................................................100.0mL
Wolfe’s vitamin solution..........................................................10.0mL
Trace elements solution SL-10 ..................................................1.0mL
pH 6.9 ± 0.2 at 25°C
Wolfe’s Vitamin Solution:
Composition per liter:
Pyridoxine·HCl........................................................................10.0mg
p-Aminobenzoic acid.................................................................5.0mg
Lipoic acid .................................................................................5.0mg
Nicotinic acid.............................................................................5.0mg
Riboflavin ..................................................................................5.0mg
Thiamine·HCl ............................................................................5.0mg
Calcium  DL -pantothenate...........................................................5.0mg
Biotin .........................................................................................2.0mg
Folic acid....................................................................................2.0mg
Vitamin B 12 ................................................................................0.1mg
Preparation of Wolfe’s Vitamin Solution:  Add components to
distilled/deionized water and bring volume to 1.0L. Mix thoroughly.
Trace Elements Solution SL-10:
Composition per liter:
FeCl 2 ·4H 2 O...................................................................................1.5g
CoCl 2 ·6H 2 O...........................................................................190.0mg
MnCl 2 ·4H 2 O ..........................................................................100.0mg
ZnCl 2 ........................................................................................70.0mg
Na 2 MoO 4 ·2H 2 O.......................................................................36.0mg
NiCl 2 ·6H 2 O..............................................................................24.0mg
H 3 BO 3 ........................................................................................6.0mg
CuCl 2 ·2H 2 O...............................................................................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 under
100% CO 2 . Add components to distilled/deionized water and bring
volume to 1.0L. Mix thoroughly. Gently heat and bring to boiling. Con-
tinue boiling for 3 min. Cool to room temperature while sparging with
100% CO 2 . Anaerobically distribute into tubes. Autoclave for 15 min
at 15 psi pressure–121°C. Adjust pH to 6.9.
Use: For the cultivation of Neocallimastix frontalis.