​Ekho Lake Strains Medium (DSMZ Medium 621a)

[所属分类:培养基配方] [发布时间:2020-7-3] [发布人:] [阅读次数:] [返回]

山东拓普生物工程有限公司

Shandong Tuopu Biol-Engineering Co.,Ltd

Ekho Lake Strains Medium
(DSMZ Medium 621a)

培养基配方
Composition per liter:
Agar.....................................................................15.0g
Peptone .................................................................0.25g
Yeast extract............................................................0.25g
Artificial sea water...................................................250.0mL
Hutner's basal salts solution...........................................20.0mL
Glucose solution .......................................................10.0mL
Vitamin solution.........................................................5.0mL
pH 7.3 ± 0.2 at 25°C
Hutner’s Basal Salts Solution:
Composition per liter:
MgSO4 ·7H2O............................................................29.7g
Nitrilotriacetic acid...................................................10.0g
CaCl2 ·2H2O ..........................................................3.335g
FeSO4 ·7H2O...........................................................99.0mg
(NH4)6MoO7O24 ·4H2O...................................................9.25mg
"Metals 44"............................................................50.0mL
“Metals 44”:
Composition per 100.0mL:
ZnSO4 ·7H2O..........................................................1.095g
FeSO4 ·7H2O............................................................0.5g
Sodium EDTA............................................................0.25g
MnSO4 ·H2O...........................................................0.154g
CuSO4 ·5H2O..........................................................39.2mg
Co(NO3)2 ·6H2O ......................................................24.8mg
Na2B4O7 ·10H2O.......................................................17.7mg
Preparation of “Metals 44”: Add sodium EDTA to distilled/de-
ionized water and bring volume to 90.0mL. Mix thoroughly. Add a few
drops of concentrated H2SO4 to retard precipitation of heavy metal
ions. Add remaining components. Mix thoroughly. Bring volume to
100.0mL with distilled/deionized water.
Preparation of Hutner’s Basal Salts Solution:  Add nitrilotria-
cetic acid to 500.0mL of distilled/deionized water. Adjust pH to 6.5
with KOH. Add remaining components. Add distilled/deionized water
to 1.0L. Adjust pH to 6.8.
Artificial Sea Water:
Composition per liter:
NaCl...................................................................23.477g
MgCl2 ·6H2O............................................................4.981g
Na2SO4 ................................................................3.917g
CaCl2 ..................................................................1.12g
KCl...................................................................664.0mg
NaHCO3 ...............................................................192.0mg
H3BO3 .................................................................26.0mg
SrCl2 .................................................................24.0mg
KBr.....................................................................6.0mg
NaF.....................................................................3.0mg
Preparation of Artificial Sea Water:  Add components to dis-
tilled/deionized water and bring volume to 1.0L. Mix thoroughly. Filter
sterilize.
Vitamin Solution:
Composition per liter:
Pyridoxine-HCl.......................................................20.0mg
Riboflavin ..........................................................10.0mg
Nicotinamide.........................................................10.0mg
Thiamine-HCl·2H2O...................................................10.0mg
Ca-pantothenate......................................................10.0mg
p-Aminobenzoic acid..................................................10.0mg
Biotin ...............................................................4.0mg
Folic acid............................................................4.0mg
Vitamin B 12 .........................................................0.2mg
Preparation of Vitamin Solution:  Add components to distilled/
deionized water and bring volume to 1.0L. Mix thoroughly. Filter ster-
ilize. Store in the dark at 5°C.
Glucose Solution:
Composition per 10.0mL:
Glucose ............................................................0.25g
Preparation of Glucose Solution:  Add glucose to distilled/deion-
ized water and bring volume to 10.0mL. Mix thoroughly. Filter steril-
ize.
Preparation of Medium: Add components, except artificial sea wa-
ter, glucose solution, and vitamin solution, to distilled/deionized water
and bring volume to 735.0mL. Mix thoroughly. Adjust pH to 7.5. Auto-
clave for 20 min at 15 psi pressure–121°C. Cool to 60°C. Warm artificial
sea water to 55°C. Aseptically add 250.0mL warm artificial sea water.
Mix thoroughly. Adjust pH to 7.3. Aseptically add 10.0mL glucose solu-
tion and 5.0 mL vitamin solution. Mix thoroughly. Aseptically distribute
into sterile tubes or flasks.
Use:  For the cultivation of Nocardioides aquaticus, Antarctobacter
heliothermus, Sulfitobacter brevis, Roseovarius tolerans, Staleya gut-
tiformis, Roseovarius tolerans, Friedmanniella lacustris, and Nest-
erenkonia lacusekhoensis.