Methanosarcina BCYT Medium (DSMZ Medium 318)

[所属分类:培养基配方] [发布时间:2021-5-26] [发布人:网站管理员2] [阅读次数:] [返回]

Methanosarcina BCYT Medium
(DSMZ Medium 318)

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

Composition per liter:

KHCO 3 ..........................................................................................2.0g
NH 4 Cl ...........................................................................................1.0g
NaCl..............................................................................................0.6g
Yeast extract..................................................................................0.5g
Trypticase™..................................................................................0.5g
KH 2 PO 4 .........................................................................................0.3g
MgCl 2 ·6H 2 O .................................................................................0.1g
CaCl 2 ·2H 2 O ................................................................................0.08g
Resazurin ...................................................................................1.0mg
Cysteine solution .....................................................................10.0mL
Na 2 S·9H 2 O solution.................................................................10.0mL
Trace elements solution...........................................................10.0mL
Vitamin solution.......................................................................10.0mL
Methanol....................................................................................5.0mL
pH 6.8 ± 0.2 at 25°C
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.
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.
Na 2 S·9H 2 O Solution :
Composition per 10.0mL:
Na 2 S·9H 2 O....................................................................................0.3g
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.
Cysteine Solution:
Composition per 10.0mL:
L -Cysteine·HCl·H 2 O .....................................................................0.3g
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.
Preparation of Medium: Add components, except methanol, vita-
min solution, cysteine solution, and Na 2 S·9H 2 O solution, to distilled/de-
ionized water and bring volume to 985.0mL. Mix thoroughly. Sparge with
100% CO 2 . Autoclave for 15 min at 15 psi pressure–121°C. Cool to 25°C
while sparging with 100% CO 2 . Aseptically and anaerobically add 5.0mL
filter sterilized methanol, 10.0mL vitamin solution, 10.0mL cysteine solu-
tion, and 10.0mL of sterile Na 2 S·9H 2 O solution. Mix thoroughly. Asepti-
cally and anaerobically distribute into sterile tubes or flasks.
Use:  For the cultivation of Methanosarcina spp.