Acetonema Medium

[所属分类:培养基配方] [发布时间:2019-9-18] [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

Acetonema Medium
Composition per liter:
Betaine·H 2 O..................................................................................6.7g
NaHCO 3 ........................................................................................4.0g
NaCl............................................................................................2.25g
Yeast extract..................................................................................2.0g
Pancreatic digest of casein............................................................2.0g
NH 4 Cl ...........................................................................................0.5g
MgSO 4 ·7H 2 O............................................................................... 0.5g
K 2 HPO 4 .....................................................................................0.348g
CaCl 2 ·2H 2 O.................................................................................0.25g
KH 2 PO 4 .....................................................................................0.227g
FeSO 4 ·7H 2 O...............................................................................2.0mg
Resazurin ...................................................................................1.0mg
NaHSeO 3 ...................................................................................26.3μg
Glucose solution ......................................................................50.0mL
Vitamin solution.......................................................................10.0mL
Reducing agent solution...........................................................10.0mL
Trace elements solution SL-10 ..................................................1.0mL
pH 7.0 ± 0.2 at 25°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.
Glucose Solution:
Composition per 50.0mL:
D- Glucose......................................................................................5.0g
Preparation of Glucose Solution: Add glucose to distilled/deion-
ized water and bring volume to 50.0mL. Mix thoroughly. Dispense so-
lution anaerobically under 100% N 2 gas. Autoclave for 15 min at 15 psi
pressure–121°C.
Reducing Agent Solution:
Composition per 10.0mL:
Dithiothreitol.............................................................................0.154g
Preparation of Reducing Agent Solution : Add 10.0mL of dis-
tilled/deionized water to a flask. Boil under N 2 gas for 1 minute. Cool
to room temperature. Add dithiothreitol and dissolve. Autoclave for 10
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 thoroughly.
Preparation of Medium: Add components, except NaHCO 3 , glucose
solution, and reducing agent solution, to distilled/deionized water and
bring volume to 940.0mL. Gently heat and bring to boiling. Continue boil-
ing for 3 min. Cool to room temperature under 80% N 2 + 20% CO 2 . Add
NaHCO 3 and bring pH to 7.0 by gassing. Distribute anaerobically under
80% N 2 + 20% CO 2 into tubes or flasks. Autoclave for 15 min at 15 psi
pressure–121°C. Prior to inoculation of cultures, aseptically and anaerobi-
cally add 0.1mL of sterile reducing agent solution and 0.5mL of sterile glu-
cose solution to each tube containing 9.4mL of sterile basal medium.
Use:  For the cultivation and maintenance of Acetonema longum and
Clostridium mayombei.