Pelobacter Medium with Gallic Acid

[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 with Gallic Acid

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

Composition per liter:

Solution A..............................................................................950.0mL
Solution B................................................................................25.0mL
Solution C................................................................................25.0mL
pH 7.2 ± 0.2 at 25°C
Solution A:
Composition per liter:
NaCl............................................................................................20.0g
MgCl 2 ·6H 2 O................................................................................3.65g
NaHCO 3 ........................................................................................2.5g
KCl................................................................................................0.5g
NH 4 Cl .........................................................................................0.25g
KH 2 PO 4 .........................................................................................0.2g
CaCl 2 ·2H 2 O.................................................................................0.15g
Resazurin ...................................................................................0.5mg
Modified Wolfe’s mineral solution..........................................10.0mL
Wolfe’s vitamin solution..........................................................10.0mL
Preparation of Solution A: Prepare and dispense solution under
80% N 2 + 20% CO 2 . Add components, except NaHCO 3 , to distilled/deion-
ized water and bring volume to 950.0mL. Mix thoroughly. Gently heat and
bring to boiling. Continue boiling for 3 min. Cool to room temperature
while sparging with 80% N 2 + 20% CO 2 . Add NaHCO 3 . Mix thoroughly.
Anaerobically distribute 9.5mL volumes into anaerobic tubes. Autoclave
for 15 min at 15 psi pressure–121°C.
Modified Wolfe’s Mineral Solution:
Composition per liter:
MgSO 4 ·7H 2 O................................................................................3.0g
Nitrilotriacetic acid.......................................................................1.5g
NaCl..............................................................................................1.0g
MnSO 4 ·H 2 O..................................................................................0.5g
CaCl 2 .............................................................................................0.1g
CoCl 2 ·6H 2 O ..................................................................................0.1g
FeSO 4 ·7H 2 O..................................................................................0.1g
ZnSO 4 ·7H 2 O.................................................................................0.1g
AlK(SO 4 ) 2 ·12H 2 O.......................................................................0.01g
CuSO 4 ·5H 2 O...............................................................................0.01g
H 3 BO 3 .........................................................................................0.01g
Na 2 MoO 4 ·2H 2 O ..........................................................................0.01g
Na 2 SeO 3 ......................................................................................0.01g
NaWO 4 ·2H 2 O..............................................................................0.01g
NiC1 2 ·6H 2 O................................................................................0.01g
Preparation of Modified Wolfe’s Mineral Solution: Add
nitrilotriacetic acid to 500.0mL of distilled/deionized water. Adjust pH
to 6.5 with KOH. Add remaining components one at a time. Add dis-
tilled/deionized water to 1.0L. Adjust pH to 6.8.
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.
Solution B:
Composition per 25.0mL:
Gallic acid...................................................................................0.85g
Preparation of Solution B: Prepare solution B immediately prior
to use. Add gallic acid to distilled/deionized water and bring volume to
25.0mL. Mix thoroughly. Rapidly adjust pH to 6.5. Filter sterilize.
Sparge with 100% N 2 .
Solution C:
Composition per 25.0mL:
Na 2 S·9H 2 O....................................................................................0.4g
Preparation of Solution C: Add Na 2 S·9H 2 O to distilled/deionized
water and bring volume to 25.0mL. Mix thoroughly. Sparge with 100%
N 2 . Autoclave for 15 min at 15 psi pressure–121°C.
Preparation of Medium: Aseptically and anaerobically add
0.25mL of sterile solution B and 0.25mL of sterile solution C to each
tube containing 9.5mL of sterile solution A. Mix thoroughly. Adjust
pH to 7.2.
Use: For the cultivation of Pelobacter acidigallici and Pelobacter
massiliensis.