Metal Acetate Agar

[所属分类:培养基配方] [发布时间:2021-5-21] [发布人:网站管理员2] [阅读次数:] [返回]

Metal Acetate Agar

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

Composition per liter:

Agar............................................................................................15.0g
Sodium acetate..............................................................................2.0g
Beijerinck's solution ................................................................50.0mL
Phosphate buffer solution........................................................50.0mL
Trace elements solution.............................................................1.0mL
pH 6.8 ± 0.2 at 25°C
Beijerinck's Solution:
Composition per liter:
NH 4 Cl .........................................................................................10.0g
MgSO 4 ·7H 2 O................................................................................0.4g
CaCl 2 ·2H 2 O ..................................................................................0.2g
Preparation of Beijerinck’s Solution: Add CaCl 2 ·2H 2 O to dis-
tilled/deionized water and bring volume to 500.0mL. Mix thoroughly.
Add the remaining components to distilled/deionized water and bring
volume to 500.0mL in a separate flask. Combine the two solutions.
Phosphate Buffer Solution:
Composition per liter:
K 2 HPO 4 .......................................................................................28.8g
KH 2 PO 4 .......................................................................................14.4g
Preparation of Phosphate Buffer Solution: Add components to
distilled/deionized water and bring volume to 1.0L. Mix thoroughly.
Adjust pH to 6.8. Autoclave for 15 min at 15 psi pressure–121°C.
Trace Elements Solution:
Composition per liter:
EDTA..........................................................................................50.0g
H 3 BO 3 solution......................................................................200.0mL
ZnSO 4 ·7H 2 O solution ............................................................100.0mL
CoCl 2 ·6H 2 O solution................................................................50.0mL
CuSO 4 ·5H 2 O solution..............................................................50.0mL
FeSO 4 ·7H 2 O solution...............................................................50.0mL
MnCl 2 ·4H 2 O solution...............................................................50.0mL
(NH 4 ) 6 Mo 7 O 24 ·4H 2 O solution .................................................50.0mL
H 3 BO 3 Solution:
Composition per 200.0mL:
H 3 BO 3 .........................................................................................11.4g
Preparation of H 3 BO 3 Solution: Add H 3 BO 3 to distilled/deion-
ized water and bring volume to 200.0mL. Mix thoroughly.
ZnSO 4 ·7H 2 O Solution:
Composition per 100.0mL:
ZnSO 4 ·7H 2 O...............................................................................22.0g
Preparation of ZnSO 4 ·7H 2 O Solution: Add ZnSO 4 ·7H 2 O to dis-
tilled/deionized water and bring volume to 100.0mL. Mix thoroughly.
MnCl 2 ·4H 2 O Solution:
Composition per 50.0mL:
MnCl 2 ·4H 2 O................................................................................5.06g
Preparation of MnCl 2 ·4H 2 O Solution : Add MnCl 2 ·4H 2 O to dis-
tilled/deionized water and bring volume to 50.0mL. Mix thoroughly.
FeSO 4 ·7H 2 O Solution:
Composition per 50.0mL:
FeSO 4 ·7H 2 O................................................................................4.99g
Preparation of FeSO 4 ·7H 2 O Solution: Add FeSO 4 ·7H 2 O to dis-
tilled/deionized water and bring volume to 50.0mL. Mix thoroughly.
CoCl 2 ·6H 2 O Solution:
Composition per 50.0mL:
CoCl 2 ·6H 2 O ................................................................................1.61g
Preparation of CoCl 2 ·6H 2 O Solution: Add CoCl 2 ·6H 2 O to dis-
tilled/deionized water and bring volume to 50.0mL. Mix thoroughly.
CuSO 4 ·5H 2 O Solution:
Composition per 50.0mL:
CuSO 4 ·5H 2 O...............................................................................1.57g
Preparation of CuSO 4 ·5H 2 O Solution: Add CuSO 4 ·5H 2 O to
distilled/deionized water and bring volume to 50.0mL. Mix thorough-
ly.
(NH 4 ) 6 Mo 7 O 24 ·4H 2 O Solution:
Composition per 50.0mL:
(NH 4 ) 6 Mo 7 O 24 ·4H 2 O....................................................................1.1g
Preparation of (NH 4 ) 6 Mo 7 O 24 ·4H 2 O Solution: Add 1.1 g of
(NH 4 ) 6 Mo 7 O 24 ·4H 2 O to distilled/deionized water and bring volume to
50.0mL. Mix thoroughly.
Preparation of Trace Elements Solution:  Add EDTA to dis-
tilled/deionized water and bring volume to 250.0mL. Mix thoroughly.
Gently heat and bring to boiling. Continue boiling until dissolved. Add
200.0mL of H 3 BO 3 solution, 100.0mL of ZnSO 4 ·7H 2 O solution,
50.0mL of MnCl 2 ·4H 2 O solution, 50.0mL of FeSO 4 ·7H 2 O solution,
50.0mL of CoCl 2 ·6H 2 O solution, 50.0mL of CuSO 4 ·5H 2 O solution,
and 50.0mL of (NH 4 ) 6 Mo 7 O 24 ·4H 2 O solution. Gently heat and bring to
boiling. Cool to 70°C. Adjust pH to 6.8 with hot (70°C) 20% KOH so-
lution. Add distilled/deionized water and bring volume to 1.0L. Allow
solution to stand in a 2.0L cotton-stoppered flask at room temperature
until the solution turns purple (approximately 2 weeks). Filter using
two layers of Whatman #1 filter paper. Filter until clear.
Preparation of Medium: Add components, except phosphate buf-
fer solution, to distilled/deionized water and bring volume to 950.0mL.
Mix thoroughly. Gently heat and bring to boiling. Autoclave for 15 min
at 15 psi pressure–121°C. Cool to 50°–55°C. Aseptically add 50.0mL
of sterile phosphate solution. Mix thoroughly. Pour into sterile Petri
dishes or distribute into sterile tubes.
Use: For the cultivation and maintenance of Chlamydomonas rein-
hardtii.