AE Sporulation Medium, Modified

[所属分类:培养基配方] [发布时间:2019-9-23] [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

AE Sporulation Medium, Modified
Composition per 1079.2mL:
Polypeptone™ ............................................................................10.0g
Yeast extract................................................................................10.0g
Na 2 HPO 4 ....................................................................................4.36g
Ammonium acetate.......................................................................1.5g
KH 2 PO 4 ......................................................................................0.25g
MgSO 4 ·7H 2 O................................................................................0.2g
Raffinose solution....................................................................39.6mL
Na 2 CO 3 solution ......................................................................13.2mL
CoCl 2 ·6H 2 O solution...............................................................13.2mL
Sodium ascorbate solution.......................................................13.2mL
pH 7.8 ± 0.1 at 25°C
Raffinose Solution:
Composition per 100.0mL:
Raffinose.....................................................................................10.0g
Preparation of Raffinose Solution: Add raffinose to distilled/de-
ionized water and bring volume to 100.0mL. Mix thoroughly. Filter
sterilize.
Na 2 CO 3 Solution:
Composition per 100.0mL:
Na 2 CO 3 .........................................................................................7.0g
Preparation of Na 2 CO 3 Solution: Add Na 2 CO 3 to distilled/deion-
ized water and bring volume to 100.0mL. Mix thoroughly. Filter ster-
ilize.
CoCl 2 ·6H 2 O Solution:
Composition per 100.0mL:
CoCl 2 ·6H 2 O................................................................................0.32g
Preparation of CoCl 2 ·6H 2 O Solution: Add CoCl 2 ·6H 2 O to dis-
tilled/deionized water and bring volume to 100.0mL. Mix thoroughly.
Filter sterilize.
Sodium Ascorbate Solution:
Composition per 100.0mL:
Sodium ascorbate..........................................................................1.5g
Preparation of Sodium Ascorbate Solution: Add sodium ascor-
bate to distilled/deionized water and bring volume to 100.0mL. Mix
thoroughly. Filter sterilize. Use freshly prepared solution.
Preparation of Medium: Add components—except raffinose solu-
tion, Na 2 CO 3 solution, CoCl 2 ·6H 2 O solution, and sodium ascorbate so-
lution—to distilled/deionized water and bring volume to 1.0L. Mix
thoroughly. Adjust pH to 7.5 using 2M sodium carbonate solution. Dis-
tribute into tubes in 15.0mL volumes. Autoclave for 15 min at 15 psi
pressure–121°C. Aseptically add 0.6mL of sterile raffinose solution,
0.2mL of sterile Na 2 CO 3 solution, and 0.2mL of sterile CoCl 2 ·6H 2 O
solution to each tube. Mix thoroughly. Prior to inoculation, steam me-
dium for 10 min. Cool to 25°C. Aseptically add 0.2mL of sterile sodi-
um ascorbate solution to each tube.
Use: For the cultivation and sporulation of Clostridium perfringens.