Pyrococcus/Staphylothermus Medium

[所属分类:培养基配方] [发布时间:2021-7-30] [发布人:网站管理员2] [阅读次数:] [返回]

Pyrococcus/Staphylothermus
Medium

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

Composition per 1010.0mL:

Sulfur, powdered.........................................................................30.0g
NaCl..........................................................................................13.85g
Peptone .........................................................................................5.0g
MgSO 4 ·7H 2 O................................................................................3.5g
MgCl 2 ·6H 2 O ...............................................................................2.75g
NiCl 2 ·6H 2 O...................................................................................2.0g
Yeast extract..................................................................................1.0g
CaCl 2 ·2H 2 O ................................................................................0.75g
KH 2 PO 4 .........................................................................................0.5g
KCl............................................................................................0.325g
NaBr............................................................................................0.05g
H 3 BO 3 .......................................................................................0.015g
(NH 4 ) 2 SO 4 ...............................................................................10.0mg
SrCl 2 ·6H 2 O ................................................................................7.5mg
Citric acid...................................................................................5.0mg
Resazurin ...................................................................................1.0mg
KI.............................................................................................0.05mg
Trace elements solution...........................................................10.0mL
Na 2 S·9H 2 O solution.................................................................10.0mL
pH 6.5 ± 0.2 at 25°C
Trace Elements Solution:
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
KAI(SO 4 ) 2 ·12H 2 O ......................................................................0.02g
CuSO 4 ·5H 2 O...............................................................................0.01g
H 3 BO 3 .........................................................................................0.01g
Na 2 MoO 4 ·2H 2 O......................................................................... 0.01g
Na 2 SeO 3 ·5H 2 O...........................................................................0.3mg
Preparation of Trace Elements Solution: Add nitrilotriacetic
acid to 500.0mL of distilled/deionized water. Adjust pH to 6.5 with
KOH. Add remaining components. Adjust pH to 7.0. Add distilled/de-
ionized water to 1.0L.
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.
Preparation of Medium: Prepare and dispense medium under
100% N 2 . Add components, except Na 2 S·9H 2 O solution, to distilled/
deionized water and bring volume to 1.0L. Mix thoroughly. Gently
heat and bring to boiling. Continue boiling for 5 min. Cool to room
temperature while sparging with 100% N 2 . Bring pH to 6.5 using 10N
H 2 SO 4 . Anaerobically distribute into tubes or flasks. Autoclave for 15
min at 15 psi pressure–121°C. Immediately prior to inoculation, add
0.1mL of sterile Na 2 S·9H 2 O solution to each 10.0mL of medium.
Check that final pH is 6.5.
Use: For the cultivation and maintenance of Pyrococcus furiosus,
Pyrococcus woesei, and Staphylothermus marinus.