CM3 Medium

[所属分类:培养基配方] [发布时间:2020-5-19] [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

CM3 Medium

培养基配方
Composition per 1040.0mL:
K2HPO4 ·3H2O.................................................................2.9g
Yeast extract.................................................................2.0g
KH2PO4 .......................................................................1.5g
(NH4)2SO4 ....................................................................1.3g
FeSO4 ·7H2O.................................................................1.25g
CaCl2 ·2H2O.................................................................0.75g
L -Cysteine·HCl..............................................................0.5g
MgCl2 ·6H2O..................................................................0.2g
Resazurin ...................................................................1.0mg
Glucose solution ...........................................................50.0mL
Na2CO3 solution.............................................................40.0mL
Na2S·9H2O solution.........................................................10.0mL
Trace elements solution SL-10 ...............................................1.0mL
pH 6.0–7.0 at 25°C
Glucose Solution:
Composition per 50.0mL:
D- Glucose.....................................................................6.0g
Preparation of Glucose Solution: Add glucose to distilled/deion-
ized water and bring volume to 50.0mL. Mix thoroughly. Sparge under
100% N2 gas for 3 min. Filter sterilize.
Na2CO3 Solution:
Composition per 40.0mL:
Na2CO3 ........................................................................2.0g
Preparation of Na2CO3 Solution:  Add Na2CO3 to distilled/deion-
ized water and bring volume to 40.0mL. Mix thoroughly. Sparge with
100% CO2 . Autoclave for 15 min at 15 psi pressure–121°C.
Na2S·9H2O Solution:
Composition per 10.0mL:
Na2S·9H2O.....................................................................0.5g
Preparation of Na2S·9H2O Solution:  Add Na2S·9H2O to dis-
tilled/deionized water and bring volume to 10.0mL. Mix thoroughly.
Sparge with 100% N2 . Autoclave for 15 min at 15 psi pressure–121°C.
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 Trace Elements Solution SL-10: Add FeCl2 ·4H2O
to 10.0mL of HCl solution. Mix thoroughly. Add distilled/deionized
water and bring volume to 1.0L. Add remaining components. Mix thor-
oughly.
Preparation of Medium: Add components, except glucose solu-
tion and Na2CO3 solution, to distilled/deionized water and bring vol-
ume to 1.0L. Mix thoroughly. Adjust pH to 6.0 with 6N HCl. Sparge
with 100% N2 . Anaerobically distribute 10.0mL volumes into screw-
capped tubes. Aseptically and anaerobically add to each tube contain-
ing 10.0mL of medium, 0.4mL of sterile Na2CO3 solution, 0.5mL of
sterile glucose solution, and 0.5mL of sterile Na2S·9H2O solution. Af-
ter addition of the Na2CO3 solution, the pH of the medium should be
6.0–7.0.
Use:  For the cultivation and maintenance of Clostridium aldrichii,
Clostridium celerecrescens, Clostridium cellulolyticum, Clostridium
lentocellum, Clostridium populeti, and Clostridium thermopalmarium.