FWM Medium

[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]

FWM Medium

培养基配方

Composition per 1013.0mL:
Solution A..............................................................................940.0mL
Solution E (NaHCO 3 solution)................................................50.0mL
Solution F (Substrate solution)................................................10.0mL
Solution G (Na 2 S·9H 2 O solution)............................................10.0mL
Solution B (Trace elements solution SL-10) .............................1.0mL
Solution C (Seven vitamin solution)..........................................1.0mL
Solution D (Selenite-tungstate solution)....................................1.0mL
pH 7.2–7.4 at 25°C
Solution A:
Composition per 940.0mL:
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
Preparation of Solution A: Prepare and dispense under 80% N 2 + 20% CO 2 . Add components to distilled/deionized water and bring volume to 940.0mL. Mix thoroughly. Autoclave for 15 min at 15 psi pres-
sure–121°C.
Solution B (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 Solution B (Trace Elements Solution SL-10):
Prepare and dispense under 100% N 2 . 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. Autoclave for 15 min at 15 psi pressure–121°C.
Solution C (Seven Vitamin Solution):
Composition per liter:
Pyridoxine·HCl......................................................................300.0mg
Nicotinic acid.........................................................................200.0mg
Thiamine·HCl ........................................................................200.0mg
Calcium  DL -pantothenate.......................................................100.0mg
Cyanocobalamine...................................................................100.0mg
p-Aminobenzoic acid...............................................................80.0mg
D (+)-Biotin...............................................................................20.0mg
Preparation of Solution C (Seven Vitamin Solution): Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Filter sterilize. Sparge with 100% N 2 .
Solution D (Selenite-Tungstate Solution):
Composition per liter:
NaOH............................................................................................0.5g
Na 2 WO 4 ·2H 2 O...........................................................................4.0mg
Na 2 SeO 3 ·5H 2 O...........................................................................3.0mg
Preparation of Solution D (Selenite-Tungstate Solution):
Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Filter sterilize. Sparge with 100% N 2 .
Solution E (NaHCO 3 Solution):
Composition per 50.0mL:
NaHCO 3 ........................................................................................2.5g
Preparation of Solution E (NaHCO 3 Solution):  Add NaHCO 3 to distilled/deionized water and bring volume to 50.0mL. Mix thoroughly. Gas under 100% N 2 . Autoclave for 15 min at 15 psi pressure– 121°C.
Solution F (Substrate Solution):
Composition per 10.0mL:
Xylan or xylose.............................................................................2.0g
Preparation of Solution F (Substrate Solution):  Add 2.0g of xylan or 2.0g of xylose to distilled/deionized water and bring volume to 10.0mL. Mix thoroughly. Gas under 100% N 2 . Autoclave for 15 min at 15 psi pressure–121°C.
Solution G (Na 2 S·9H 2 O Solution):
Composition per 10.0mL:
Na 2 S·9H 2 O....................................................................................0.3g
Preparation of Solution G (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. Gas under 100% N 2 . Autoclave for 15 min at 15 psi pressure–121°C.
Preparation of Medium: To 940.0mL of sterile solution A, aseptically and anaerobically add 1.0mL of sterile solution B, 1.0mL of sterile solution C, 1.0mL of sterile solution D, 50.0mL of sterile solution E, 10.0mL of sterile solution F, and 10.0mL of sterile solution G. Mix thoroughly. Aseptically and anaerobically distribute into sterile tubes or flasks.
Use:  For the cultivation and maintenance of Desulfococcus biacutus