Methanopyrus Medium

[所属分类:培养基配方] [发布时间:2021-5-26] [发布人:网站管理员2] [阅读次数:] [返回]

Methanopyrus 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..........................................................................................11.80g
MgCl 2 ·6H 2 O................................................................................4.50g
MgSO 4 ·7H 2 O..............................................................................1.75g
Na 2 SO 4 ........................................................................................0.81g
CaCl 2 ·2H 2 O ................................................................................0.78g
KCl..............................................................................................0.30g
KH 2 PO 4 .......................................................................................0.09g
K 2 HPO 4 ·3H 2 O ............................................................................0.07g
Na 2 WO 4 ·2H 2 O...........................................................................2.0mg
(NH 4 ) 2 Fe (SO 4 ) 2 ·6H 2 O ..............................................................2.0mg
(NH 4 ) 2 Ni(SO 4 ) 2 .........................................................................2.0mg
Resazurin ...................................................................................0.2mg
Marine trace elements..............................................................10.0mL
Trace elements solution...........................................................10.0mL
Vitamin solution.......................................................................10.0mL
NaHCO 3 solution.....................................................................10.0mL
Na 2 S·9H 2 O solution.................................................................10.0mL
pH 6.5 ± 0.2 at 25°C
Marine Trace Elements:
Composition per liter:
NaBr..............................................................................................4.0g
SrCl 2 ·6H 2 O ...................................................................................1.8g
H 3 BO 3 ...........................................................................................1.3g
Sodium silicate.......................................................................100.0mg
NaF ..........................................................................................60.0mg
KNO 3 .......................................................................................40.0mg
KI.............................................................................................1.25mg
Na 2 HPO 4 ·3H 2 O........................................................................0.25mg
Preparation of Marine Trace Elements:  Add components to
distilled/deionized water and bring volume to 1.0L. Mix thoroughly.
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
KAl(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.
Vitamin Solution:
Composition per liter:
Pyridoxine·HCl........................................................................10.0mg
Calcium  DL -pantothenate...........................................................5.0mg
Lipoic acid.................................................................................5.0mg
Nicotinic acid.............................................................................5.0mg
p-Aminobenzoic acid.................................................................5.0mg
Riboflavin..................................................................................5.0mg
Thiamine·HCl ............................................................................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. Filter ster-
ilize. Sparge with 80% H 2 + 20% CO 2 .
NaHCO 3 Solution:
Composition per 10.0mL:
NaHCO 3 ........................................................................................0.5g
Preparation of NaHCO 3 Solution:  Add NaHCO 3 to distilled/de-
ionized water and bring volume to 10.0mL. Mix thoroughly. Sparge
with 80% N 2 + 20% CO 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.
Preparation of Medium: Prepare and dispense medium under 80%
H 2 + 20% CO 2 . Add components, except vitamin solution, NaHCO 3
solution, and Na 2 S·9H 2 O solution, to distilled/deionized water and
bring volume to 970.0mL. Mix thoroughly. Anaerobically distribute
into tubes or flasks fitted with butyl rubber stoppers. Autoclave for 15
min at 15 psi pressure–121°C. Anaerobically add 10.0mL of sterile vi-
tamin solution, 10.0mL of sterile NaHCO 3 solution, and 10.0mL of
sterile Na 2 S·9H 2 O solution to each liter of medium or, using a syringe,
inject the appropriate amount of sterile vitamin solution, sterile
NaHCO 3 solution, and sterile Na 2 S·9H 2 O solution into individual
tubes containing medium. Check that final pH of medium is 6.5.
Use: For the cultivation and maintenance of Methanopyrus kandleri.