Anaerobacter Medium

[所属分类:培养基配方] [发布时间:2019-10-10] [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

Anaerobacter Medium
Composition per liter:
Yeast extract..................................................................................1.0g
CaCl 2 ·2H 2 O ................................................................................0.33g
KCl..............................................................................................0.33g
KH 2 PO 4 .......................................................................................0.33g
MgCl 2 ·6H 2 O ...............................................................................0.33g
NH 4 Cl .........................................................................................0.33g
Resazurin ...................................................................................1.0mg
NaHCO 3 solution.....................................................................30.0mL
Sucrose solution.......................................................................10.0mL
L -Cysteine·HCl solution ..........................................................10.0mL
Na 2 S·9H 2 O solution.................................................................10.0mL
Trace elements solution SL-10..................................................1.0mL
pH 7.0 ± 0.2 at 25°C
NaHCO 3 Solution:
Composition per 30.0mL:
NaHCO 3 ........................................................................................1.5g
Preparation of NaHCO 3 Solution : Add NaHCO 3 to distilled/de-
ionized water and bring volume to 30.0mL. Mix thoroughly. Sparge
under 80% N 2 + 20% CO 2 for 3 min. Autoclave for 15 min at 15 psi
pressure–121°C.
Sucrose Solution:
Composition per 100.0mL:
Sucrose........................................................................................20.0g
Preparation of Sucrose Solution : Add sucrose to distilled/deion-
ized water and bring volume to 100.0mL. Mix thoroughly. Sparge with
100% N 2 . Autoclave under 100% N 2 for 15 min at 15 psi pressure–
121°C.
L -Cysteine·HCl Solution:
Composition per 10.0mL:
L -Cysteine·HCl .............................................................................0.3g
Preparation of  L -Cysteine·HCl Solution : Add  L -cysteine·HCl to
distilled/deionized water and bring volume to 10.0mL. Mix thorough-
ly. Sparge with 100% N 2 . Autoclave under 100% N 2 for 15 min at 15
psi pressure–121°C.
Na 2 S·9H 2 O Solution:
Composition per 10.0mL:
Na 2 S·9H 2 O....................................................................................0.3g
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.
Sparge with 100% N 2 . Autoclave under 100% N 2 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.
Preparation of Medium: Add components, except NaHCO 3 solu-
tion, sucrose solution,  L -cysteine·HCl solution, and Na 2 S·9H 2 O solu-
tion, to distilled/deionized water and bring volume to 940.0mL. Mix
thoroughly. Sparge under 100% N 2 for 3–4 min. Autoclave under
100% N 2 for 15 min at 15 psi pressure–121°C. Aseptically and anaer-
obically add the sterile NaHCO 3 solution, sucrose solution,  L -
cysteine·HCl solution, and Na 2 S·9H 2 O solution. Mix thoroughly. Final
pH of the medium should be 7.0.
Use:  For the cultivation and maintenance of Anaerobacter polyen-
dosporus.