FWN Medium, Modified with Xylan

[所属分类:培养基配方] [发布时间:2021-3-5] [发布人: ] [阅读次数:] [返回]

FWN Medium, Modified with Xylan

培养基配方

Composition per 1020.0mL:
Solution A..............................................................................950.0mL
Solution C (NaHCO 3 solution)................................................50.0mL
Solution D (Na 2 S·9H 2 O solution)............................................10.0mL
Solution B (Wolfe’s vitamin solution).....................................10.0mL
Solution A:
Composition per 950.0mL:
Xylan.............................................................................................2.0g
NaCl..............................................................................................1.0g
KCl................................................................................................0.5g
MgCl 2 ·6H 2 O..................................................................................0.4g
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
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 distilled/deionized water to 1.0L. Adjust pH to 6.8.
Preparation of Solution A: Prepare and dispense under 80% N 2 + 20% CO 2 . Add components to distilled/deionized water and bring volume to 950.0mL. Mix thoroughly. Autoclave for 15 min at 15 psi pressure–121°C.
Solution B (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 Solution B (Wolfe’s Vitamin Solution):  Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Sparge with 100% N 2 . Filter sterilize.
Solution C (NaHCO 3 Solution):
Composition per 50.0mL:
NaHCO 3 ........................................................................................2.5g
Preparation of Solution C (NaHCO 3 Solution):  Add NaHCO 3 to distilled/deionized water and bring volume to 50.0mL. Mix thoroughly. Sparge with 100% N 2 . Autoclave for 15 min at 15 psi pressure–121°C.
Solution D (Na 2 S·9H 2 O Solution):
Composition per 10.0mL:
Na 2 S·9H 2 O....................................................................................0.3g
Preparation of Solution D (Na 2 S·9H 2 O Solution):  Add Na 2 S·9H 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. Before use, neutralize to pH 7.0 with sterile HCl. 

Preparation of Medium: To 950.0mL of sterile solution A, aseptically and anaerobically add 10.0mL of sterile solution B, 50.0mL of sterile solution C, and 10.0mL of sterile solution D. Mix thoroughly. Aseptically and anaerobically distribute into sterile tubes or flasks under 100% N 2 .
Use: For the cultivation of Cytophaga xylanolytica.