Natranaerobius Medium (DSMZ Medium 1095)

[所属分类:培养基配方] [发布时间:2021-6-16] [发布人:网站管理员2] [阅读次数:] [返回]

Natranaerobius Medium
(DSMZ Medium 1095)

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..........................................................................................100.0g
Yeast extract................................................................................10.0g
Tryptone......................................................................................10.0g
NH 4 Cl ..........................................................................................0.5g
KH 2 PO 4 .........................................................................................0.2g
MgCl 2 ·6H 2 O..................................................................................0.1g
Sucrose solution.......................................................................10.0mL
Vitamin solution.......................................................................10.0mL
Cysteine solution......................................................................10.0mL
Bicarbonate solution................................................................10.0mL
Carbonate solution...................................................................10.0mL
Trace elements solution SL-10 ..................................................1.0mL
pH 8.5 ± 0.2 at 25°C
Vitamin Solution:
Composition per liter:
Pyridoxine-HCl........................................................................10.0mg
Thiamine-HCl·2H 2 O..................................................................5.0mg
Riboflavin ..................................................................................5.0mg
Nicotinic acid.............................................................................5.0mg
D -Ca-pantothenate......................................................................5.0mg
p-Aminobenzoic acid.................................................................5.0mg
Lipoic acid .................................................................................5.0mg
Biotin .........................................................................................2.0mg
Folic acid....................................................................................2.0mg
Vitamin B 12 ................................................................................0.1mg
Preparation of Vitamin Solution:  Add components to distilled/
deionized water and bring volume to 1.0L. Mix thoroughly. Sparge
with 80% H 2 + 20% CO 2 . Filter sterilize.
Trace Elements Solution SL-10:
Composition per liter:
MgSO 4 ·7H 2 O................................................................................3.0g
Nitrilotriacetic acid.......................................................................1.5g
NaCl..............................................................................................1.0g
MnSO 4 ·2H 2 O................................................................................0.5g
CoSO 4 ·7H 2 O...............................................................................0.18g
ZnSO 4 ·7H 2 O...............................................................................0.18g
CaCl 2 ·2H 2 O ..................................................................................0.1g
FeSO 4 ·7H 2 O..................................................................................0.1g
NiCl 2 ·6H 2 O...............................................................................0.025g
KAl(SO 4 ) 2 ·12H 2 O.......................................................................0.02g
H 3 BO 3 .........................................................................................0.01g
Na 2 MoO 4 ·4H 2 O..........................................................................0.01g
CuSO 4 ·5H 2 O...............................................................................0.01g
Na 2 SeO 3 ·5H 2 O...........................................................................0.3mg
Preparation of Trace Elements Solution SL-10:  Add nitrilotri-
acetic acid to 500.0mL of distilled/deionized water. Dissolve by adjust-
ing pH to 6.5 with KOH. Add remaining components. Add distilled/
deionized water to 1.0L. Mix thoroughly. Adjust pH to 7.0.
Sucrose Solution :
Composition per 10.0mL:
Sucrose..........................................................................................5.0g
Preparation of Sucrose Solution:  Add sucrose to distilled/deion-
ized water and bring volume to 10.0mL. Mix thoroughly. Filter steril-
ize.
Carbonate Solution :
Composition per 10.0mL:
Na 2 CO 3 .......................................................................................68.0g
Preparation of Carbonate Solution:  Add Na 2 CO 3 to distilled/de-
ionized water and bring volume to 10.0mL. Mix thoroughly. Filter ster-
ilize.
Bicarbonate Solution :
Composition per 10.0mL:
NaHCO 3 ......................................................................................38.0g
Preparation of Bicarbonate Solution:  Add NaHCO 3 to distilled/
deionized water and bring volume to 10.0mL. Mix thoroughly. Filter
sterilize.
Cysteine Solution:
Composition per 10.0mL:
L- Cysteine-HCl·2H 2 O.................................................................0.07g
Preparation of Cysteine Solution: Add  L- Cysteine-HCl·2H 2 O to
to distilled/deionized water and bring volume to 10.0mL. Mix thor-
oughly. Sparge with 100% N 2 . Filter sterilize.
Preparation of Medium:  Add components, except carbonate, bi-
carbonate, cysteine, sucrose, and vitamin solutions, to distilled/deion-
ized water and bring volume to 950.0mL. Mix thoroughly. Sparge wtih
100% N 2 gas. Gently heat and bring to boiling. Boil for 1 min. Cool to
room temperature while sparging with 100% N 2 . Add cysteine, carbon-
ate, and bicarbonate solutions. Adjust pH to 8.5. Autoclave for 15 min at
15 psi pressure–121°C. Cool to room temperature. Aseptically add the
vitamins and sucrose solutions. Aseptically and anoxically dispense
into cuture vessels.
Use: For the cultivation of Natranaerobius spp.