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Medium for Roseospira

[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 for Roseospira

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............................................................................................20.0g
MgCl 2 ·6H 2 O .................................................................................1.0g
MgSO 4 · 7H 2 O..............................................................................0.25g
NH 4 Cl ...........................................................................................0.5g
Yeast extract..................................................................................0.5g
KH 2 PO 4 .........................................................................................0.3g
CaCl 2 ·2H 2 O.................................................................................0.05g
NaHCO 3  solution .....................................................................10.0mL
Acetate solution .......................................................................10.0mL
Succinate solution....................................................................10.0mL
Na 2 S·9H 2 O solution.................................................................10.0mL
Trace elements solution SL-12 ..................................................1.0mL
Vitamin V7 solution...................................................................1.0mL
pH 6.9 ± 0.2 at 25°C
Acetate Solution:
Composition per 10.0mL:
Sodium acetate............................................................................0.41g
Preparation of Acetate Solution:  Add sodium acetate to distilled/
deionized water and bring volume to 10.0mL. Mix thoroughly. Sparge
with 20% CO 2 + 80% N 2 . Filter sterilize.
Succinate Solution:
Composition per 10.0mL:
Sodium succinate........................................................................0.85g
Preparation of Succinate Solution:  Add sodium succinate to dis-
tilled/deionized water and bring volume to 10.0mL. Mix thoroughly.
Sparge with 20% CO 2 + 80% N 2 . Filter sterilize.
NaHCO 3 Solution :
Composition per 10.0mL:
NaHCO 3 ........................................................................................1.5g
Preparation of NaHCO 3 Solution:  Add NaHCO 3 to distilled/de-
ionized water and bring volume to 10.0mL. Mix thoroughly. Sparge
with 20% CO 2 + 80% N 2 . Filter sterilize.
Vitamin Solution V7:
Composition per liter:
Pyridoxine-HCl........................................................................50.0mg
Nicotinic acid...........................................................................20.0mg
Vitamin B 12 ..............................................................................20.0mg
Thiamine-HCl·2H 2 O................................................................10.0mg
p-Aminobenzoic acid...............................................................10.0mg
D -Ca-pantothenate......................................................................5.0mg
Biotin .........................................................................................2.0mg
Preparation of Vitamin Solution V7:  Add components to dis-
tilled/deionized water and bring volume to 1.0L. Mix thoroughly. Filter
sterilize.
Trace Elements Solution SL-12:
Composition per liter:
FeSO 4 ·7H 2 O..................................................................................1.1g
H 3 BO 3 ...........................................................................................0.3g
CoCl 2 ·6H 2 O ................................................................................0.19g
MnCl 2 ·2H 2 O................................................................................0.05g
ZnCl 2 ........................................................................................42.0mg
NiCl 2 ·6H 2 O..............................................................................24.0mg
Na 2 MoO 4 ·4H 2 O .......................................................................18.0mg
CuCl 2 ·2H 2 O ...............................................................................2.0mg
Preparation of Trace Elements Solution Sl-12:  Add compo-
nents to distilled/deionized water and bring volume to 1.0L. Mix thor-
oughly. Filter sterilize.
Na 2 S·9H 2 O Solution :
Composition per 10.0mL:
Na 2 S·9H 2 O....................................................................................0.2g
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.
Autoclave under 100% N 2 for 15 min at 15 psi pressure–121°C. Cool
to room temperature.
Preparation of Medium: Add components, except sulfide, bicar-
bonate, vitamin, acetate, and succinate solutions, to distilled/deionized
water and bring volume to 970.0mL. Mix thoroughly. Gently heat
while stirring and bring to boiling. Boil for 1 min. Cool to room tem-
perature while sparging with 90% N 2 + 10% CO 2 gas. Autoclave for 15
min at 15 psi pressure–121°C. Aseptically add bicarbonate and vitamin
solutions. Mix thoroughly. Adjust pH to 6.9. Distribute into sterile
50mL screw-capped bottles. Add sulfide and organic acetate and suc-
cinate substrates.
Use: For the cultivation of  Roseospira navarrensis.
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