Ilyobacter polytropus Medium

[所属分类:培养基配方] [发布时间:2021-4-20] [发布人:李佳颐] [阅读次数:] [返回]

Ilyobacter polytropus Medium

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

Composition per 1011.0mL:

Solution A.....................................................................................1.0L
Solution B................................................................................10.0mL
Vitamin solution.........................................................................1.0mL
pH 7.2–7.4 at 25°C
Solution A:
Composition per liter:
NaHCO 3 ........................................................................................4.5g
Na 2 SO 4 ........................................................................................2.84g
Sodium 3-hydroxybutyrate...........................................................1.3g
NaCl............................................................................................1.17g
Yeast extract..................................................................................1.0g
MgCl 2 ·6H 2 O .................................................................................0.4g
KCl................................................................................................0.3g
NH 4 Cl .........................................................................................0.27g
KH 2 PO 4 .........................................................................................0.2g
CaCl 2 ·2H 2 O ................................................................................0.15g
Resazurin...................................................................................0.5mg
Trace elements solution.............................................................1.0mL
Preparation of Solution A: Add components, except NaHCO 3 and
vitamin solution, and bring volume to 1.0L. Mix thoroughly. Gently
heat and bring to boiling. Continue boiling for 3 to 4 min. Allow to cool
to room temperature while gassing under O 2 -free 80% N 2 + 20% CO 2 .
Add NaHCO 3 and continue gassing with O 2 -free 80% N 2 + 20% CO 2
until pH reaches 6.9–7.1. Seal the flask under 80% N 2 + 20% CO 2 . Au-
toclave for 15 min at 15 psi pressure–121°C.
Trace Elements Solution:
Composition per liter:
FeCl 2 ·4H 2 O...................................................................................1.5g
CoCl 2 ·6H 2 O...........................................................................120.0mg
MnCl 2 ·4H 2 O ..........................................................................100.0mg
ZnCl 2 ........................................................................................68.0mg
H 3 BO 3 ......................................................................................62.0mg
Na 2 MoO 4 ·2H 2 O...................................................................... 24.0mg
NiCl 2 ·6H 2 O..............................................................................24.0mg
CuCl 2 ·2H 2 O.............................................................................17.0mg
HCl (25% solution)..................................................................10.0mL
Preparation of Trace Elements Solution:  Add FeCl 2 ·4H 2 O to
10.0mL of HCl solution. Mix thoroughly. Add distilled/deionized wa-
ter and bring volume to 1.0L. Add remaining components. Mix thor-
oughly. Gas under 100% N 2 .
Vitamin Solution:
Composition per 100.0mL:
Thiamine·HCl ..........................................................................10.0mg
p-Aminobenzoic acid.................................................................4.0mg
D (+)-Biotin.................................................................................1.0mg
Preparation of Vitamin Solution:  Add components to distilled/
deionized water and bring volume to 100.0mL. Mix thoroughly. Filter
sterilize. Gas under 100% N 2 .
Solution B:
Composition per 10.0mL:
Na 2 S·9H 2 O..................................................................................0.36g
Preparation of Solution B:  Add Na 2 S·9H 2 O to distilled/deionized
water and bring volume to 10.0mL. Mix thoroughly. Gas under 100%
N 2 . Autoclave for 15 min at 15 psi pressure–121°C.
Preparation of Medium: To 1.0L of sterile solution A, add 10.0mL
of sterile solution B and 1.0mL of sterile vitamin solution. Mix thor-
oughly. Adjust final pH to 7.2–7.4.
Use:  For the cultivation and maintenance of Ilyobacter polytropus.