Dialister Medium (DSMZ Medium 1107)

[所属分类:培养基配方] [发布时间:2020-6-16] [发布人:] [阅读次数:] [返回]

山东拓普生物工程有限公司

Shandong Tuopu Biol-Engineering Co.,Ltd

Dialister Medium
(DSMZ Medium 1107)

培养基配方
Composition per liter:
Trypticase..................................................................17.0g
Yeast extract................................................................3.0g
NaCl.........................................................................3.0g
KNO3 ........................................................................3.0g
Lactate solution...........................................................10.0mL
Hemin solution.............................................................10.0mL
Glucose soltuion ..........................................................10.0mL
L -Cysteine solution........................................................5.0mL
DTT solution................................................................5.0mL
Vitamin K 1 solution .......................................................5.0mL
Formiate solution..........................................................50.0mL
Fumarate solution .........................................................50.0mL
Horse blood, sterile.......................................................50.0mL
pH 7.0 ± 0.2 at 25°C
L -Cystine Solution:
Composition per 5.0mL:
L- Cystine ..................................................................0.25g
NaOH (1N solution)...........................................................5.0mL
Preparation of  L -Cystine Solution: Add  L- cystine to 5.0mL of
NaOH solution. Mix thoroughly. Sparge with 100% N2 . Filter sterilize.
Vitamin K 1 Solution:
Composition per 100.0mL:
Vitamin K 1 .................................................................0.5g
Ethanol.....................................................................99.0mL
Preparation of Vitamin K 1 Solution:  Add vitamin K 1 to 99.0mL
of absolute ethanol. Mix thoroughly. Sparge with 100% N2 . Filter ster-
ilize. Store refrigerated.
Hemin Solution:
Composition per 100.0mL:
Hemin.....................................................................50.0mg
NaOH (1N solution)........................................................20.0mL
Preparation of Hemin Solution:  Add hemin to 20.0mL of 1N
NaOH solution. Mix thoroughly. Bring volume to 100.0mL with dis-
tilled/deionized water. Sparge with 100% N2 . Filter sterilize.
Lactate Solution:
Composition per 10.0mL:
Sodium lactate.............................................................5.0g
Preparation of Lactate Solution:  Add sodium lactate to distilled/
deionized water and bring volume to 10.0mL. Mix thoroughly. Sparge
with 100% N2 . Filter sterilize.
Glucose Solution:
Composition per 10.0mL:
Glucose ...................................................................4.0g
Preparation of Glucose Solution:  Add glucose to distilled/deion-
ized water and bring volume to 10.0mL. Mix thoroughly. Sparge with
100% N2 . Filter sterilize.
Formiate Solution:
Composition per 10.0mL:
Na-formiate...............................................................0.6g
Preparation of Formiate Solution:  Add Na-formiate to distilled/
deionized water and bring volume to 10.0mL. Mix thoroughly. Sparge
with 100% N2 . Filter sterilize.
Fumarate Solution:
Composition per 10.0mL:
Na-fumarate...............................................................0.6g
Preparation of Fumarate Solution:  Add Na-fumarate to dis-
tilled/deionized water and bring volume to 10.0mL. Mix thoroughly.
Sparge with 100% N2 . Filter sterilize.
DTT Solution:
Composition per 10.0mL:
DTT.......................................................................0.15g
Preparation of DTT Solution:  Add DTT to distilled/deionized
water and bring volume to 10.0mL. Mix thoroughly. Sparge with 100%
N2 . Filter sterilize.
NaHCO3 Solution :
Composition per 10.0mL:
NaHCO3 ....................................................................1.0g
Preparation of NaHCO3 Solution:  Add NaHCO3 to distilled/de-
ionized water and bring volume to 10.0mL. Mix thoroughly. Sparge
with 20% CO2 + 80% H2 . Filter sterilize.
Preparation of Medium: Add components, except horse blood,
lactate, hemin, glucose, cysteine, DTT, Vitamin K1, formiate, and fu-
marate solutions, to distilled/deionized water and bring volume to
810.0mL. Mix thoroughly. Adjust pH to 7.8. Gently heat while stirring
and bring to boiling. Distribute into tubes or bottles. Autoclave for 15
min at 15 psi pressure–121°C. Aseptically and anaerobically add horse
blood, lactate, hemin, glucose, cysteine, DTT, vitamin K 1 , formiate,
and fumarate solutions. Incubate in oxygen-free, 5–10% CO2 contain-
ing atmosphere.
Use: For the cultivation of Dialister fulvus, Dialister vulgaris, and
other Dialister spp.