LuPhet1 Medium (DSMZ Medium 298e)

[所属分类:培养基配方] [发布时间:2021-5-8] [发布人:网站管理员2] [阅读次数:] [返回]

LuPhet1 Medium
(DSMZ Medium 298e)

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

Composition per liter:

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 ...................................................................................1.0mg
NaHCO 3  solution .....................................................................10.0mL
Sodium lactate solution............................................................10.0mL
Na 2 S·9H 2 O solution.................................................................10.0mL
Yeast extract solution...............................................................10.0mL
Seven vitamin solution...............................................................1.0mL
Selenite-tungstate solution.........................................................1.0mL
Trace elements solution SL-10 ..................................................1.0mL
pH 7.2 ± 0.2 at 25°C
Na 2 S·9H 2 O Solution :
Composition per 10.0mL:
Na 2 S·9H 2 O..................................................................................0.36g
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.
Autoclave under 100% N 2 for 15 min at 15 psi pressure–121°C. Cool
to room temperature.
NaHCO 3 Solution:
Composition per 10.0mL:
NaHCO 3 ........................................................................................5.0g
Preparation of NaHCO 3 Solution:  Add NaHCO 3 to distilled/de-
ionized water and bring volume to 10.0mL. Mix thoroughly. Sparge
with 80% N 2 + 20% CO 2 . Filter sterilize.
Sodium Lactate Solution:
Composition per 10.0mL:
Sodium lactate...............................................................................1.5g
Preparation of Sodium Lactate Solution: Add sodium lactate to
distilled/deionized water and bring volume to 10.0mL. Mix thorough-
ly. Sparge with 100% N 2 . Filter sterilize.
Yeast Extract Solution:
Composition per 10.0mL:
Yeast extract..................................................................................1.0g
Preparation of Yeast Extract Solution: Add yeast extract to dis-
tilled/deionized water and bring volume to 10.0mL. Mix thoroughly.
Sparge with 100% N 2 . Filter sterilize.
Seven Vitamin Solution:
Composition per liter:
Pyridoxine hydrochloride ......................................................300.0mg
Thiamine-HCl·2H 2 O..............................................................200.0mg
Nicotinic acid.........................................................................200.0mg
Vitamin B 12 ............................................................................100.0mg
Calcium pantothenate ............................................................100.0mg
p-Aminobenzoic acid...............................................................80.0mg
D (+)-Biotin...............................................................................20.0mg
Preparation of Seven Vitamin Solution: Add components to dis-
tilled/deionized water and bring volume to 1.0L. Sparge with 100% N 2 .
Mix thoroughly. Filter sterilize.
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 Trace Elements Solution SL-10: 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 thor-
oughly. Sparge with 80% N 2 + 20% CO 2 . Autoclave for 15 min at 15
psi pressure–121°C.
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 Selenite-Tungstate Solution: Add components
to distilled/deionized water and bring volume to 1.0L. Mix thoroughly.
Sparge with 100% N 2 . Filter sterilize.
Preparation of Medium: Prepare and dispense medium under 80%
N 2 + 20% CO 2 gas atmosphere. Add components, except NaHCO 3  solu-
tion, sodium lactate solution, Na 2 S·9H 2 O solution, seven vitamin solu-
tion, selenite-tungstate solution, and trace elements solution SL-10, to
distilled/deionized water and bring volume to 957.0mL. Mix thoroughly.
Adjust pH to 7.2. Sparge with 80% N 2 + 20% CO 2 . Autoclave for 15 min
at 15 psi pressure–121°C. Aseptically and anaerobically add 10.0mL
NaHCO 3  solution, 10.0mL Na 2 S·9H 2 O solution, 10.0mL sodium lac-
tate solution, 1.0mL seven vitamin solution, 1.0mL selenite- tungstate
solution, and 1.0mL trace elements solution SL-10. Mix thoroughly.
Aseptically and anaerobically distribute into sterile tubes or bottles. Af-
ter inoculation, flush and repressurize the gas head space of culture bot-
tles with sterile 80% N 2 + 20% CO 2 to 1 bar overpressure.
Use:  For the cultivation of Acetobacterium sp.