Pelobacter Medium

[所属分类:培养基配方] [发布时间:2021-7-1] [发布人:网站管理员2] [阅读次数:] [返回]

Pelobacter Medium

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

Composition per liter:

KHCO 3 ..........................................................................................4.5g
NH 4 Cl ...........................................................................................1.0g
NaCl..............................................................................................0.6g
Trypticase™..................................................................................0.5g
Yeast extract..................................................................................0.5g
KH 2 PO 4 .........................................................................................0.3g
MgCl 2 ·6H 2 O..................................................................................0.1g
CaCl 2 ·2H 2 O.................................................................................0.08g
Resazurin ...................................................................................1.0mg
Trace elements solution ...........................................................10.0mL
Vitamin solution.......................................................................10.0mL
Sodium gallate solution ...........................................................10.0mL
L -Cysteine·HCl·H 2 O solution ..................................................10.0mL
Na 2 S·9H 2 O solution.................................................................10.0mL
pH 7.3 ± 0.2 at 25°C
Trace Elements Solution:
Composition per liter:
Nitrilotriacetic acid.....................................................................12.8g
FeCl 3 ·6H 2 O.................................................................................1.35g
NaCl..............................................................................................1.0g
NiCl 2 ·6H 2 O.................................................................................0.12g
CaCl 2 ·2H 2 O.................................................................................0.10g
MnCl 2 ·4H 2 O ...............................................................................0.10g
ZnCl 2 ...........................................................................................0.10g
Na 2 SeO 3 ·5H 2 O..........................................................................0.026g
CuCl 2 ·2H 2 O..............................................................................0.025g
CoCl 2 ·6H 2 O..............................................................................0.024g
Na 2 MoO 4 ·2H 2 O........................................................................0.024g
H 3 BO 3 .........................................................................................0.01g
Preparation of Trace Elements Solution: Add nitrilotriacetic
acid to 500.0mL of distilled/deionized water. Adjust pH to 6.5 with
KOH. Add remaining components. Adjust pH to 7.0. Add distilled/de-
ionized water to 1.0L.
Vitamin Solution:
Composition per liter:
Biotin.........................................................................................2.0mg
Folic acid ...................................................................................2.0mg
Pyridoxine·HCl........................................................................10.0mg
Thiamine·HCl............................................................................5.0mg
Riboflavin..................................................................................5.0mg
Nicotinic acid.............................................................................5.0mg
Calcium  DL -pantothenate...........................................................5.0mg
Vitamin B 12 ................................................................................0.1mg
p-Aminobenzoic acid.................................................................5.0mg
Lipoic acid.................................................................................5.0mg
Preparation of Vitamin Solution : Add components to distilled/
deionized water and bring volume to 1.0L. Mix thoroughly. Filter ster-
ilize. Sparge with 80% N 2 + 20% CO 2 .
Sodium Gallate Solution:
Composition per 10.0mL:
Gallic acid...................................................................................1.88g
NaOH (1M solution)............................................................... variable
Preparation of Sodium Gallate Solution:  Add gallic acid to dis-
tilled/deionized water and bring volume to 8.0mL. Mix thoroughly.
Add sufficient NaOH solution to bring pH to 7.3. Filter sterilize.
Sparge with 80% N 2 + 20% CO 2 .
L -Cysteine·HCl·H 2 O Solution:
Composition per 10.0mL:
L -Cysteine·HCl·H 2 O.....................................................................0.3g
Preparation of  L -Cysteine·HCl·H 2 O Solution: Add  L -cyste-
ine·HCl·H 2 O to distilled/deionized water and bring volume to 10.0mL.
Mix thoroughly. Sparge with 100% N 2 . Autoclave 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 for 15 min at 15 psi pressure–121°C.
Preparation of Medium: Prepare and dispense medium under 80%
N 2 + 20% CO 2 . Add components, except sodium gallate solution,
NaHCO 3  solution, Na 2 S·9H 2 O solution, and vitamin solution, to dis-
tilled/deionized water and bring volume to 960.0mL. Mix thoroughly.
Sparge with 80% N 2 + 20% CO 2 . Autoclave for 15 min at 15 psi pres-
sure–121°C. Aseptically and anaerobically add 10.0mL of sterile
NaHCO 3  solution, 10.0mL of sterile Na 2 S·9H 2 O solution, 10.0mL of
sterile sodium gallate solution, and 10.0mL of sterile vitamin solution
or, using a syringe, inject the appropriate amount of sterile NaHCO 3
solution, sterile Na 2 S·9H 2 O solution, sterile sodium gallate solution,
and sterile vitamin solution into individual tubes containing medium.
Use: For the cultivation and maintenance of Pelobacter acidigallici.