MB Medium (DSMZ Medium 924)

[所属分类:培养基配方] [发布时间:2021-5-17] [发布人:网站管理员2] [阅读次数:] [返回]

MB Medium
(DSMZ Medium 924)

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............................................................................................10.0g
NaHCO 3 ........................................................................................4.0g
Yeast extract..................................................................................2.0g
Trypticase™..................................................................................2.0g
NH 4 Cl ...........................................................................................1.0g
MgCl 2 ·6H 2 O .................................................................................1.0g
KCl................................................................................................0.5g
CaCl 2 ·2H 2 O ..................................................................................0.4g
K 2 HPO 4 .........................................................................................0.4g
Resazurin ...................................................................................0.5mg
Sodium formate solution..........................................................50.0mL
Na 2 S·9H 2 O solution.................................................................10.0mL
Cysteine-HCl·H 2 O solution .....................................................10.0mL
Vitamin solution.......................................................................10.0mL
Trace elements solution ...........................................................10.0mL
Selenite-tungstate solution.........................................................1.0mL
pH 7.2 ± 0.2 at 25°C
Cysteine Solution:
Composition per 10.0mL:
L -Cysteine·HCl·H 2 O ...................................................................0.25g
Preparation of Cysteine Solution:  Add  L -cysteine·HCl·H 2 O to
distilled/deionized water and bring volume to 10.0mL. Mix thorough-
ly. Sparge with 100% N 2 . Autoclave for 15 min at 15 psi pressure–
121°C.
Sodium Formate Solution:
Composition per 50.0mL:
Na-formate....................................................................................6.8g
Preparation of Sodium Formate Solution: Add sodium formate
to distilled/deionized water and bring volume to 50.0mL. Mix thor-
oughly. Sparge with 100% N 2 . Filter sterilize.
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.
Na 2 S·9H 2 O Solution:
Composition per 10.0mL:
Na 2 S·9H 2 O..................................................................................0.25g
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. Sparge with N 2 .
Autoclave for 15 min at 15 psi pressure–121°C. Cool to 25°C. Store an-
aerobically.
Trace Elements Solution:
Composition per liter:
MgSO 4 ·7H 2 O................................................................................3.0g
Nitrilotriacetic acid.......................................................................1.5g
NaCl..............................................................................................1.0g
MnSO 4 ·2H 2 O................................................................................0.5g
CoSO 4 ·7H 2 O...............................................................................0.18g
ZnSO 4 ·7H 2 O...............................................................................0.18g
CaCl 2 ·2H 2 O...................................................................................0.1g
FeSO 4 ·7H 2 O..................................................................................0.1g
NiCl 2 ·6H 2 O...............................................................................0.025g
KAl(SO 4 ) 2 ·12H 2 O.......................................................................0.02g
H 3 BO 3 .........................................................................................0.01g
Na 2 MoO 4 ·4H 2 O ..........................................................................0.01g
CuSO 4 ·5H 2 O...............................................................................0.01g
Na 2 SeO 3 ·5H 2 O...........................................................................0.3mg
Preparation of Trace Elements Solution:  Add nitrilotriacetic
acid to 500.0mL of distilled/deionized water. Dissolve by adjusting pH
to 6.5 with KOH. Add remaining components. Add distilled/deionized
water to 1.0L. Mix thoroughly.
Vitamin Solution:
Composition per liter:
Pyridoxine-HCl........................................................................10.0mg
Thiamine-HCl·2H 2 O..................................................................5.0mg
Riboflavin..................................................................................5.0mg
Nicotinic acid.............................................................................5.0mg
D -Ca-pantothenate .....................................................................5.0mg
p-Aminobenzoic acid.................................................................5.0mg
Lipoic acid.................................................................................5.0mg
Biotin.........................................................................................2.0mg
Folic acid ...................................................................................2.0mg
Vitamin B 12 ................................................................................0.1mg
Preparation of Vitamin Solution:  Add components to distilled/
deionized water and bring volume to 1.0L. Mix thoroughly. Sparge
with 80% H 2 + 20% CO 2 . Filter sterilize.
Preparation of Medium: Prepare and dispense medium under an
oxygen-free 80% N 2 + 20% CO 2  gas mixture. Add components, except
sodium formate solution, NaHCO 3 , cysteine solution, and Na 2 S·9H 2 O
solution, to distilled/deionized water and bring volume to 920.0mL.
Mix thoroughly. Gently heat and bring to boiling. Boil for 3 min. Cool
to 25°C while sparging with 80% N 2 + 20% CO 2 . Add solid NaHCO 3 .
Mix thoroughly. Adjust pH to 6.8–7.0. Distribute into tubes or bottles.
Autoclave for 15 min at 15 psi pressure–121°C. Aseptically and anaer-
obically add per liter 50.0mL sterile sodium formate solution, 10.0mL
of sterile cysteine solution, and 10.0mL of sterile Na 2 S·9H 2 O solution.
Mix thoroughly. The final pH should be 7.2.
Use:  For the cultivation of Methanocalculus taiwanensis, Methanococ-
cus voltae (Methanococcus voltaei), and Methanofollis aquaemaris.