Petrotoga Medium

[所属分类:培养基配方] [发布时间:2021-7-5] [发布人:网站管理员2] [阅读次数:] [返回]

Petrotoga 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:

NaCl............................................................................................18.0g
MgSO 4 ·7H 2 O..............................................................................3.45g
MgCl 2 ·7H 2 O ...............................................................................2.75g
NaHCO 3 ........................................................................................1.0g
L- Cysteine·HCl·H 2 O .....................................................................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 ·7H 2 O...............................................................2.0mg
Resazurin...................................................................................1.0mg
Glucose solution......................................................................50.0mL
Trace elements solution SL-6 ..................................................10.0mL
Pancreatic digest of casein solution.........................................10.0mL
Yeast extract solution...............................................................10.0mL
Na 2 S·9H 2 O solution.................................................................10.0mL
Wolfe’s vitamin solution..........................................................10.0mL
pH 6.5–6.7 at 25°C
Glucose Solution:
Composition per 50.0mL:
D -Glucose......................................................................................5.0g
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.
Trace Elements Solution SL-6:
Composition per liter:
H 3 BO 3 ...........................................................................................0.3g
CoCl 2 ·6H 2 O ..................................................................................0.2g
ZnSO 4 ·7H 2 O.................................................................................0.1g
MnCl 2 ·4H 2 O................................................................................0.03g
Na 2 MoO 4 ·H 2 O ............................................................................0.03g
NiCl 2 ·6H 2 O.................................................................................0.02g
CuCl 2 · 2H 2 O.................................................................................0.01g
Preparation of Trace Elements Solution SL-6: Add components
to distilled/deionized water and bring volume to 1.0L. Mix thoroughly.
Adjust pH to 3.4.
Pancreatic Digest of Casein Solution:
Composition per 10.0mL:
Pancreatic digest of casein............................................................1.0g
Preparation of Pancreatic Digest of Casein Solution:  Add
pancreatic digest of casein to distilled/deionized water and bring vol-
ume to 10.0mL. Mix thoroughly. Sparge with 100% N 2 . Autoclave for
15 min at 15 psi pressure–121°C.
Yeast Extract Solution:
Composition per 10.0mL:
Yeast extract..................................................................................1.0g
Preparation of Yeast Extract Solution:  Add yeast extract to dis-
tilled/deionized water and bring volume to 10.0mL Mix thoroughly.
Sparge with 100% N 2 . Autoclave for 15 min at 15 psi pressure–121°C.
Na 2 S·9H 2 O Solution:
Composition per 10.0mL:
Na 2 S·9H 2 O....................................................................................0.5g
Preparation of Na 2 S·9H 2 O Solution:  Add Na 2 S·9H 2 O to dis-
tilled/deionized water and bring volume to 10.0mL. 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
Thiamine·HCl ............................................................................5.0mg
Riboflavin ..................................................................................5.0mg
Nicotinic acid.............................................................................5.0mg
Calcium pantothenate ................................................................5.0mg
p-Aminobenzoic acid.................................................................5.0mg
Thioctic acid ..............................................................................5.0mg
Biotin .........................................................................................2.0mg
Folic acid....................................................................................2.0mg
Cyanocobalamin .....................................................................100.0μg
Preparation of Wolfe’s Vitamin Solution: Add components to
distilled/deionized water and bring volume to 1.0L. Mix thoroughly.
Filter sterilize. Sparge with 100% N 2 .
Preparation of Medium:  Prepare and dispense medium under 80%
N 2 + 20% CO 2 . Add components, except NaHCO 3 , glucose solution,
pancreatic digest of casein solution, yeast extract solution, Na 2 S·9H 2 O
solution, and Wolfe’s vitamin solution, to distilled/deionized water and
bring volume to 910.0mL. Mix thoroughly. Adjust pH to 6.5–6.7. Gen-
tly heat and bring to boiling. Continue boiling for 3 min. Cool to room
temperature while sparging with 80% N 2 + 20% CO 2 . Add NaHCO 3 .
Adjust pH to 6.5–6.7. Anaerobically distribute 9.1mL volumes into an-
aerobic tubes. Autoclave for 15 min at 15 psi pressure–121°C. Asepti-
cally add 0.5mL of sterile glucose solution, 0.1mL of sterile pancreatic
digest of casein solution, 0.1mL of sterile yeast extract solution, 0.1mL
of sterile Na 2 S·9H 2 O solution, and 0.1mL of sterile Wolfe’s vitamin so-
lution to each tube. Mix thoroughly.
Use: For the cultivation of Petrotoga miotherma.