Medium VTY

[所属分类:培养基配方] [发布时间:2021-5-20] [发布人:网站管理员2] [阅读次数:] [返回]

Medium VTY

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

Composition per 100.0mL:

Peptone .........................................................................................1.0g
Noble agar.....................................................................................0.7g
Yeast extract..................................................................................0.5g
L- Cysteine·HCl·H 2 O .....................................................................0.1g
Salts A......................................................................................20.0mL
Salts B......................................................................................20.0mL
Glucose solution........................................................................5.0mL
NaHCO 3 (5% solution)..............................................................1.0mL
Hemin solution...........................................................................1.0mL
Volatile fatty acid solution.......................................................0.31mL
Resazurin (0.1% solution) .........................................................0.1mL
pH 7.2 ± 0.2 at 25°C
NaHCO 3 Solution:
Composition per 10.0mL:
NaHCO 3 ........................................................................................0.5g
Preparation of NaHCO 3 Solution:  Add NaHCO 3 to distilled/de-
ionized water and bring volume to 10.0mL. Mix thoroughly. Sparge
with 100% N 2 . Filter sterilize.
Glucose Solution:
Composition per 10.0mL:
Glucose.........................................................................................1.0g
Preparation of Glucose Solution:  Add glucose to distilled/deion-
ized water and bring volume to 10.0mL. Mix thoroughly. Filter steril-
ize.
Salts A:
Composition per liter:
CaCl 2 ·2H 2 O...................................................................................0.6g
MgSO 4 ........................................................................................0.45g
Preparation of Salts A: Add components to distilled/deionized wa-
ter and bring volume to 1.0L. Mix thoroughly.
Salts B:
Composition per liter:
NaCl..............................................................................................4.5g
(NH 4 ) 2 SO 4 .....................................................................................4.5g
Potassium phosphate
buffer (0.05M, pH 7.4) ...........................................................1.0L
Preparation of Salts B: Add NaCl and (NH 4 ) 2 SO 4 to 1.0L of 0.05M
potassium phosphate buffer, pH 7.4. Mix thoroughly.
Hemin Solution:
Composition per liter:
Hemin............................................................................................0.5g
NaOH (0.01N solution)..............................................................1.0mL
Preparation of Hemin Solution: Add hemin to 1.0mL of 0.01N
NaOH solution. Mix thoroughly.
Volatile Fatty Acid Solution:
Composition per 31.0mL:
Acetic acid ...............................................................................17.0mL
Propionic acid............................................................................6.0mL
n-Butyric acid ............................................................................4.0mL
n-Valeric acid.............................................................................1.0mL
Isovaleric acid............................................................................1.0mL
Isobutyric acid............................................................................1.0mL
DL- α-Methylbutyric acid............................................................1.0mL
Preparation of Volatile Fatty Acid Solution: Combine compo-
nents. Mix thoroughly.
Preparation of Medium: Add components, except glucose and
NaHCO 3  solutions, to distilled/deionized water and bring volume to
94.0mL. Mix thoroughly. Adjust pH to 7.2. Gently heat and gas with
95% N 2 + 5% CO 2 until reduced. Anaerobically distribute into tubes or
flasks. Cap with rubber stoppers. Autoclave for 20 min at 15 psi pres-
sure–121°C. Cool to 50°C. Filter sterilize glucose solution and
NaHCO 3 solution separately. Aseptically and anaerobically add sterile
glucose solution and sterile NaHCO 3 solution to cooled, sterile basal
medium.
Use:  For the cultivation and maintenance of Roseburia cecicola.