ACE Medium

[所属分类:培养基配方] [发布时间:2019-9-18] [发布人:] [阅读次数:] [返回]

山东拓普生物工程有限公司

Shandong Tuopu Biol-Engineering Co.,Ltd

ACE Medium
Composition per liter:
NaCl............................................................................................18.0g
MgCl 2 ·6H 2 O .................................................................................6.0g
NaHCO 3 ........................................................................................2.0g
MgSO 4 ·7H 2 O................................................................................1.0g
Yeast extract..................................................................................1.0g
L -Cysteine·HCl .............................................................................0.5g
KCl............................................................................................0.335g
NH 4 Cl .........................................................................................0.25g
CaCl 2 ·2H 2 O ................................................................................0.14g
K 2 HPO 4 .......................................................................................0.14g
Fe(NH 4 ) 2 (SO 4 ) 2 ·6H 2 O...............................................................2.0mg
Resazurin...................................................................................1.0mg
Glucose solution......................................................................20.0mL
Wolfe’s vitamin solution..........................................................10.0mL
pH 7.6 ± 0.2 at 25°C
Glucose Solution:
Composition per 50.0mL:
D -Glucose....................................................................................10.0g
Preparation of Glucose Solution:  Add glucose to distilled/deion-
ized water and bring volume to 50.0mL. Mix thoroughly. Sparge with
100% N 2 . Autoclave for 15 min at 15 psi pressure–121°C.
Wolfe’s Vitamin Solution:
Composition per liter:
Pyridoxine·HCl........................................................................10.0mg
p-Aminobenzoic acid.................................................................5.0mg
Lipoic acid.................................................................................5.0mg
Nicotinic acid.............................................................................5.0mg
Riboflavin..................................................................................5.0mg
Thiamine·HCl............................................................................5.0mg
Calcium  DL -pantothenate...........................................................5.0mg
Biotin.........................................................................................2.0mg
Folic acid ...................................................................................2.0mg
Vitamin B 12 ................................................................................0.1mg
Preparation of Wolfe’s Vitamin Solution:  Add components to
distilled/deionized water and bring volume to 1.0L. Mix thoroughly.
Filter sterilize.
Preparation of Medium: Add components, except  L -cysteine·HCl,
NaHCO 3 , and glucose solution, to distilled/deionized water and bring
volume to 980.0mL. Mix thoroughly. Adjust pH to 7.6. Gently heat and
bring to boiling. Continue boiling for 3 min. Cool to room temperature
while sparging with O 2 -free 100% N 2 . Add  L -cysteine·HCl and
NaHCO 3.  Anaerobically distribute 9.8mL volumes into anaerobic
tubes. Autoclave for 15 min at 15 psi pressure–121°C. Aseptically and
anaerobically add 0.2mL of sterile glucose solution to each tube. Mix
thoroughly.
Use:  For the cultivation of unclassified bacterium DSM 6211 and
unclassified bacterium DSM 6226