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培養基配方
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M10 Broth
[所屬分類:培養基配方] [發布時間:2021-11-29] [發布人:網站管理員2] [閱讀次數:] [返回]
M10 Broth
山東拓普生物工程有限公司 培養基配方 //ljxinglong.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
Na2CO3 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:
K2HPO4...............................................................0.2g
Preparation of Mineral Solution I: Add
K2HPO4 to distilled/deionized water and bring vol
ume to 100.0mL. Mix thoroughly.
Mineral Solution II:
Composition per 100.0mL:
NaCl....................................................................0.4g
(NH4)2SO4...........................................................0.4g
KH2PO4...............................................................0.3g
MgSO4·7H2O ....................................................0.09g
CaCl2.................................................................0.05g
Preparation of Mineral Solution II: Add com
ponents to distilled/deionized water and bring vol
ume to 100.0mL. Mix thoroughly.
Na2CO3 Solution:
Composition per 100.0mL:
Na2CO3 ...............................................................8.0g
Preparation of Na2CO3 Solution: Add Na2CO3
to distilled/deionized water and bring volume to
100.0mL. Mix thoroughly. Sparge with 100% CO2.
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 compo
nents 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 thoroughly. Autoclave under
100% N2 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 components. Mix thoroughly. Store under
100% N2.
Preparation of Medium: Prepare and dispense
medium under 100% CO2. Add components, except
L-cysteine·HCl solution and Na2CO3 solution, to dis
tilled/deionized water and bring volume to 930.0mL.
Mix thoroughly. Sparge with 100% CO2. Adjust pH
to 6.8 with 1N NaOH. Distribute anaerobically
9.3mL volumes into Hungate tubes. Autoclave for 15
min at 15 psi pressure–121°C. Aseptically and anaer
obically add 0.2mL of sterile L-cysteine·HCl solution
and 0.5mL of sterile Na2CO3 solution. Check that fi
nal pH is 6.8. (Note: if not properly gassed with
100% CO2, the medium pH can be as high as 9.5.)
Use: For the cultivation of Enterococcus species,
Lactobacillus species and Streptococcus species.



