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
Cellobiose solution.......................................................50.0mL
Na2CO3 solution ..........................................................40.0mL
Trace elements solution SL-10..............................................1.0mL
pH 7.2 ± 0.2 at 25°C
Cellobiose Solution:
Composition per 50.0mL:
D- Cellobiose...............................................................6.0g
Preparation of Cellobiose Solution: Add cellobiose to distilled/
deionized 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.
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 cellobiose 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 0.4mL of sterile
Na2CO3 solution to each tube containing 10.0mL of medium and
0.5mL of sterile cellobiose solution to each tube. pH of the medium af-
ter the addition of Na2CO3 solution should be 7.2.
Use:  For the cultivation of Clostridium aldrichii, Clostridium celerec-
rescens, Clostridium cellulolyticum, Clostridium lentocellum, Clostrid-
ium populeti, and Clostridium thermopalmarium.