AMO.1 Medium

[所属分类:培养基配方] [发布时间:2019-10-9] [发布人:] [阅读次数:] [返回]

山东拓普生物工程有限公司

Shandong Tuopu Biol-Engineering Co.,Ltd

Composition per 1012.0mL:
Yeast extract................................................................................10.0g
NaCl..............................................................................................5.8g
N-Methylhydantoin.....................................................................5.64g
NaHCO 3 ........................................................................................4.5g
L -Serine.........................................................................................2.0g
L -Threonine...................................................................................2.0g
Pancreatic digest of casein............................................................0.5g
L -Cysteine.....................................................................................0.5g
MgCl 2 ·6H 2 O .................................................................................0.4g
KCl................................................................................................0.3g
NH 4 Cl .........................................................................................0.27g
KH 2 PO 4 .........................................................................................0.2g
CaCl 2 ·2H 2 O ................................................................................0.15g
Wolfe’s vitamin solution..........................................................10.0mL
NaHSeO 3  solution......................................................................1.0mL
Trace elements solution SL-10..................................................1.0mL
pH 8.3 ± 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.
Na 2 SeO 3 Solution:
Composition per liter:
Na 2 SeO 3 ·5H 2 O...........................................................................0.2mg
Preparation of Na 2 SeO 3 Solution:  Add Na 2 SeO 3 ·5H 2 O to dis-
tilled/deionized water and bring volume to 1.0L. 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:
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. Sparge with 100% N 2 . Autoclave for 15 min at 15 psi pressure–
121°C.
Preparation of Medium:  Prepare anaerobically .  Add components,
except NaHCO 3 and  L -cysteine, to distilled/deionized water and bring
volume to 1.0L. Sparge with 100% N 2 for 30 min. Adjust pH to 8.3
with 10N NaOH. Add NaHCO 3 and  L -cysteine (solid substances). Mix
thoroughly. Sparge with 80% N 2 + 20% CO 2 mixture. Flush the head-
space of the medium vessel with the 80% N 2 + 20% CO 2 mixture. Au-
toclave for 15 min at 15 psi pressure–121°C. Sparge with 80% N 2 +
20% CO 2 mixture for 30 min. Aseptically and anaerobically distribute
into sterile tubes or bottles.
Use:  For the cultivation of Tissierella creatinini.