ML Medium (Minimal Lactate Medium)

[所属分类:培养基配方] [发布时间:2021-6-3] [发布人:网站管理员2] [阅读次数:] [返回]

ML Medium
(Minimal Lactate Medium)

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

Composition per liter:

Sodium lactate...............................................................................5.0g
MgSO 4 ·7H 2 O................................................................................2.0g
NH 4 Cl ...........................................................................................1.0g
Na 2 SO 4 ..........................................................................................1.0g
Yeast extract..................................................................................1.0g
K 2 HPO 4 .........................................................................................0.5g
L -Cysteine.....................................................................................0.5g
CaCl 2 ·6H 2 O...................................................................................0.1g
Resazurin ...................................................................................1.0mg
NaHCO 3 solution.....................................................................25.0mL
FeSO 4 ·7H 2 O solution...............................................................25.0mL
pH 6.8 ± 0.2 at 25°C
NaHCO 3 Solution:
Composition per 25.0mL:
NaHCO 3 ........................................................................................4.0g
Preparation of NaHCO 3 Solution: Add NaHCO 3  to distilled/de-
ionized water and bring volume to 25.0mL. Mix thoroughly. Filter ster-
ilize. Gas with O 2 -free 97% N 2 + 3% H 2 . Cap with a rubber stopper.
FeSO 4 ·7H 2 O Solution:
Composition per 25.0mL:
FeSO 4 ·7H 2 O...............................................................................4.0mg
Preparation of FeSO 4 ·7H 2 O Solution: Add FeSO 4 ·7H 2 O to dis-
tilled/deionized water and bring volume to 25.0mL. Mix thoroughly.
Filter sterilize. Gas with O 2 -free 97% N 2 + 3% H 2 . Cap with a rubber
stopper.
Preparation of Medium: Add components, except NaHCO 3 solu-
tion and FeSO 4 ·7H 2 O solution, to distilled/deionized water and bring
volume to 1.0L. Gently heat and bring to boiling under O 2 -free 97% N 2
+ 3% H 2 . Adjust pH to 6.8. Continue boiling until resazurin becomes
colorless, indicating reduction. Distribute anaerobically under O 2 -free
97% N 2 + 3% H 2 into tubes in 10.0mL volumes. Cap with rubber stop-
pers. Place tubes in a press. Autoclave for 15 min at 15 psi pressure–
121°C. Cool to room temperature. Prior to inoculation, add 0.25mL of
sterile NaHCO 3 solution and 0.25mL of sterile FeSO 4 ·7H 2 O solution
to each test tube containing 10.0mL of sterile basal medium.
Use:  For the cultivation and maintenance of Desulfovibrio species.