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培养基配方
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TMBS4 Medium
[所属分类:培养基配方] [发布时间:2021-9-16] [发布人:网站管理员2] [阅读次数:] [返回]
TMBS4 Medium
山东拓普生物工程有限公司 培养基配方 http://www.topbiol.com
Composition per 1002.0mL:
Solution A ..............................................................................870.0mL
Solution C ..............................................................................100.0mL
Solution D ................................................................................10.0mL
Solution E (Vitamin solution)..................................................10.0mL
Solution F.................................................................................10.0mL
Solution B (Trace elements solution SL-10) .............................1.0mL
Solution G..................................................................................1.0mL
pH 7.1–7.4 at 25°C
Solution A:
Composition per 870.0mL:
Na2SO4.......................................................................................... 3.0g
NaCl.............................................................................................. 1.0g
KCl................................................................................................ 0.5g
MgCl2·6H2O ................................................................................. 0.4g
NH4Cl ........................................................................................... 0.3g
KH2PO4......................................................................................... 0.2g
CaCl2·2H2O ................................................................................ 0.15g
Resazurin ................................................................................... 1.0mg
Preparation of Solution A: Add components to distilled/deionized
water and bring volume to 870.0mL. Mix thoroughly. Gently heat and
bring to boiling. Continue boiling for 3–4 min. Allow to cool to room
temperature while gassing under 80% N2 + 20% CO2. Continue gas
sing until pH reaches below 6.0. Seal the flask under 80% N2 + 20%
CO2. Autoclave for 15 min at 15 psi pressure–121°C.
Solution B (Trace Elements Solution SL-10):
Composition per liter:
FeCl2·4H2O................................................................................... 1.5g
CoCl2·6H2O ........................................................................... 190.0mg
MnCl2·4H2O .......................................................................... 100.0mg
ZnCl2........................................................................................ 70.0mg
Na2MoO4·2H2O ....................................................................... 36.0mg
NiCl2·6H2O.............................................................................. 24.0mg
H3BO3 ........................................................................................ 6.0mg
CuCl2·2H2O ............................................................................... 2.0mg
HCl (25% solution)..................................................................10.0mL
Preparation of Solution B (Trace Elements Solution SL-10):
Add FeCl2·4H2O to 10.0mL of HCl solution. Mix thoroughly. Add dis
tilled/deionized water and bring volume to 1.0L. Add remaining com
ponents. Mix thoroughly. Gas under 100% N2. Autoclave for 15 min at
15 psi pressure–121°C.
Solution C:
Composition per 100.0mL:
NaHCO3........................................................................................ 5.0g
Preparation of Solution C: Add NaHCO3 to distilled/deionized
water and bring volume to 100.0mL. Mix thoroughly. Filter sterilize.
Gas under 80% N2 + 20% CO2.
Solution D:
Composition per 10.0mL:
Syringic acid ................................................................................. 0.5g
Preparation of Solution D: Add syringic acid to distilled/deion
ized water and bring volume to 10.0mL. Mix thoroughly. Adjust pH to
8 with NaOH. Gas under 100% N2. Autoclave for 15 min at 15 psi pres
sure–121°C.
Solution E (Vitamin Solution):
Composition per liter:
Pyridoxine·HCl ........................................................................ 10.0mg
Calcium DL-pantothenate........................................................... 5.0mg
Lipoic acid ................................................................................. 5.0mg
Nicotinic acid............................................................................. 5.0mg
p-Aminobenzoic acid................................................................. 5.0mg
Riboflavin .................................................................................. 5.0mg
Thiamine·HCl ............................................................................5.0mg
Biotin .........................................................................................2.0mg
Folic acid....................................................................................2.0mg
Vitamin B12 ................................................................................0.1mg
Preparation of Solution E (Vitamin Solution): Add compo
nents to distilled/deionized water and bring volume to 1.0L. Mix thor
oughly. Gas under 100% N2. Autoclave for 15 min at 15 psi pressure–
121°C.
Solution F:
Composition per 10.0mL:
Na2S·9H2O.................................................................................. 0.25g
Na2S2O4 ...................................................................................... 0.25g
Preparation of Solution F: Add components to distilled/deionized
water and bring volume to 10.0mL. Mix thoroughly. Gas under 100%
N2. Autoclave for 15 min at 15 psi pressure–121°C.
Solution G:
Composition per liter:
NaOH ............................................................................................ 0.5g
Na2WO4·2H2O...........................................................................4.0mg
Na2SeO3·5H2O...........................................................................3.0mg
Preparation of Solution G: Add components to distilled/deionized
water and bring volume to 1.0L. Mix thoroughly. Gas under 100% N2.
Autoclave for 15 min at 15 psi pressure–121°C.
Preparation of Medium: Aseptically and anaerobically combine
solution A with solution B, solution C, solution D, solution E, solution
F, and solution G, in that order. Mix thoroughly. Anaerobically distrib
ute into sterile tubes or flasks under 80% N2 + 20% CO2.
Use: For the cultivation and maintenance of Pelobacter species.