M10 Broth

[所属分类:培养基配方] [发布时间:2021-5-11] [发布人:网站管理员2] [阅读次数:] [返回]

M10 Broth

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

Composition per liter:

Pancreatic digest of casein............................................................2.0g
Yeast extract..................................................................................2.0g
Cellobiose .....................................................................................1.0g
Glucose .........................................................................................1.0g
Maltose..........................................................................................1.0g
Starch ............................................................................................1.0g
Resazurin ...................................................................................1.0mg
Mineral solution I...................................................................100.0mL
Mineral solution II.................................................................100.0mL
Na 2 CO 3  solution......................................................................50.0mL
Hemin solution.........................................................................10.0mL
L -Cysteine·HCl solution...........................................................10.0mL
Volatile fatty acid mixture..........................................................3.1mL
pH 6.8 ± 0.2 at 25°C
Mineral Solution I:
Composition per 100.0mL:
K 2 HPO 4 .........................................................................................0.2g
Preparation of Mineral Solution I : Add K 2 HPO 4 to distilled/de-
ionized water and bring volume to 100.0mL. Mix thoroughly.
Mineral Solution II:
Composition per 100.0mL:
NaCl..............................................................................................0.4g
(NH 4 ) 2 SO 4 .....................................................................................0.4g
KH 2 PO 4 .........................................................................................0.3g
MgSO 4 ·7H 2 O..............................................................................0.09g
CaCl 2 ...........................................................................................0.05g
Preparation of Mineral Solution II : Add components to dis-
tilled/deionized water and bring volume to 100.0mL. Mix thoroughly.
Na 2 CO 3 Solution:
Composition per 100.0mL:
Na 2 CO 3  .........................................................................................8.0g
Preparation of Na 2 CO 3 Solution : Add Na 2 CO 3  to distilled/deion-
ized water and bring volume to 100.0mL. Mix thoroughly. Sparge with
100% CO 2 . Autoclave for 15 min at 15 psi pressure–121°C.
Hemin Solution:
Composition per 100.0mL:
Hemin ...........................................................................................1.0g
NaOH (1N solution).................................................................10.0mL
Preparation of Hemin Solution:  Add components to 100.0mL of
distilled/deionized water. Mix thoroughly.
L -Cysteine·HCl Solution:
Composition per 10.0mL:
L -Cysteine·HCl ...........................................................................0.25g
Preparation of  L -Cysteine·HCl Solution: Add  L -cysteine·HCl to
distilled/deionized water and bring volume to 10.0mL. Mix thorough-
ly. Autoclave under 100% N 2 for 15 min at 15 psi pressure–121°C.
Volatile Fatty Acid Mixture:
Composition per 31.0mL:
Acetic acid...............................................................................17.0mL
Propionic acid............................................................................6.0mL
Butyric acid................................................................................4.0mL
DL -α-Methylbutyric acid ...........................................................1.0mL
Isobutyric acid ...........................................................................1.0mL
Isovaleric acid............................................................................1.0mL
n-Valeric acid.............................................................................1.0mL
Preparation of Volatile Fatty Acid Mixture : Combine compo-
nents. Mix thoroughly. Store under 100% N 2 .
Preparation of Medium: Prepare and dispense medium under
100% CO 2 . Add components, except  L -cysteine·HCl solution and
Na 2 CO 3  solution, to distilled/deionized water and bring volume to
930.0mL. Mix thoroughly. Sparge with 100% CO 2 . Adjust pH to 6.8
with 1N NaOH. Distribute anaerobically in 9.3mL volumes into Hun-
gate tubes. Autoclave for 15 min at 15 psi pressure–121°C. Aseptically
and anaerobically add 0.2mL of sterile  L -cysteine·HCl solution and
0.5mL of sterile Na 2 CO 3  solution. Check that final pH is 6.8.
Use: For the cultivation of Enterococcus species, Lactobacillus spe-
cies, Streptococcus species, and Vagococcus fluvialis.