M13 Verrucomicrobium Medium

[所属分类:培养基配方] [发布时间:2021-5-11] [发布人:网站管理员2] [阅读次数:] [返回]

M13 Verrucomicrobium 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:

Glucose.......................................................................................0.25g
Peptone .......................................................................................0.25g
Yeast extract................................................................................0.25g
Distilled water........................................................................670.0mL
Artificial seawater..................................................................250.0mL
Tris-HCl buffer, (0.1M solution, pH 7.5).................................50.0mL
Modified Huntner’s basal salts................................................20.0mL
Vitamin solution.......................................................................10.0mL
pH 7.5 ± 0.2 at 25°C
Artificial Seawater:
Composition per liter:
NaCl..........................................................................................23.48g
MgCl 2 ..........................................................................................4.98g
Na 2 SO 4 ........................................................................................3.92g
CaCl 2 .............................................................................................1.1g
KCl..............................................................................................0.66g
NaHCO 3 ......................................................................................0.19g
H 3 BO 3 .......................................................................................0.026g
SrCl 2 ..........................................................................................0.024g
KBr ............................................................................................6.0mg
NaF ............................................................................................3.0mg
Preparation of Artificial Seawater: Add components to distilled/
deionized water and bring volume to 1.0L. Mix thoroughly.
Modified Hutner’s Basal Salts:
Composition per liter:
MgSO 4 ·7H 2 O..............................................................................29.7g
Nitrilotriacetic acid.....................................................................10.0g
CaCl 2 ·2H 2 O.................................................................................3.34g
FeSO 4 ·7H 2 O.............................................................................99.0mg
(NH 4 ) 2 MoO 4 ............................................................................9.25mg
Metals “44”..............................................................................50.0mL
Preparation of Modified Hutner’s Basal Salts: Add nitrilotria-
cetic 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.
Metals “44”:
Composition per 100.0mL:
ZnSO 4 ·7H 2 O.................................................................................1.1g
FeSO 4 ·7H 2 O..................................................................................0.5g
EDTA..........................................................................................0.25g
MnSO 4 ·7H 2 O............................................................................0.154g
CuSO 4 ·5H 2 O...............................................................................0.04g
Co(NO 3 ) 2 ·6H 2 O ........................................................................0.025g
Na 2 B 4 O 7 ·10H 2 O........................................................................0.018g
Preparation of Metals “44”: Add components to distilled/deion-
ized water and bring volume to 100.0mL. Mix thoroughly.
Vitamin Solution:
Composition per liter:
D- Calcium pantothenate.............................................................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.
Preparation of Medium: Add components, except modified Hut-
ner’s basal salts, to distilled/deionized water and bring volume to
980.0mL. Mix thoroughly. Autoclave for 15 min at 15 psi pressure–
121°C. Cool to room temperature. Aseptically add 20.0mL of sterile
modified Hutner’s basal salts. Mix thoroughly. Aseptically distribute
into sterile tubes or flasks.
Use: For the cultivation and maintenance of Verrucomicrobium spino-
sum.