Pine and Drouhet’s Histoplasma Yeast Phase Medium

[所属分类:培养基配方] [发布时间:2021-7-8] [发布人:网站管理员2] [阅读次数:] [返回]

Pine and Drouhet’s Histoplasma
Yeast Phase 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:

Casein hydrolysate-vitamin-
glucose base.....................................................................500.0mL
Agar starch base.....................................................................500.0mL
Casein Hydrolysate-Vitamin-Glucose Base:
Composition per 500.0mL:
Glucose .......................................................................................10.0g
Citric acid....................................................................................10.0g
L -Cysteine·HCl..............................................................................1.0g
L -Asparagine.................................................................................1.0g
Glutathione, reduced.....................................................................0.5g
L -Tryptophan...............................................................................0.02g
Solution I................................................................................250.0mL
Solution II................................................................................10.0mL
Solution III...............................................................................40.0mL
Solution IV...............................................................................10.0mL
Solution V..................................................................................1.0mL
Solution VI.................................................................................0.1mL
Solution VII ...............................................................................0.1mL
Solution I:
Composition per liter:
KH 2 PO 4 .........................................................................................8.0g
(NH 4 ) 2 SO 4 .....................................................................................8.0g
MgSO 4 ·7H 2 O..............................................................................0.86g
CaCl 2 ...........................................................................................0.08g
Preparation of Solution I: Add components to distilled/deionized
water and bring volume to 1.0L. Mix thoroughly. Store at 5°C.
Solution II:
Composition per liter:
FeSO 4 ·7H 2 O..................................................................................5.7g
MnCl 2 ·6H 2 O..................................................................................0.8g
Na 2 MoO 4 ·2H 2 O ..........................................................................0.15g
HCl, concentrated ......................................................................1.0mL
Preparation of Solution II: Add the concentrated HCl to 100.0mL
of distilled/deionized water. Add FeSO 4 ·7H 2 O, MnCl 2 ·6H 2 O, and
Na 2 MoO 4 ·2H 2 O, in that order. Make sure each salt is completely dis-
solved before adding the next. Mix thoroughly. Bring volume to 1.0L
with distilled/deionized water. Mix thoroughly. Store at 5°C. Solution
is good until a red precipitate forms.
Solution III:
Composition per 100.0mL:
Acid hydrolysate of casein, vitamin free.......................................10.0g
Preparation of Solution III: Add acid hydrolysate of casein to dis-
tilled/deionized water and bring volume to 100.0mL. Mix thoroughly.
Gently heat until dissolved.
Solution IV:
Composition per liter:
Inositol..........................................................................................0.2g
Thiamine·HCl ...............................................................................0.2g
Calcium pantothenate ...................................................................0.2g
Riboflavin.....................................................................................0.2g
Nicotinamide.................................................................................0.1g
Biotin.......................................................................................10.0mg
Preparation of Solution IV: Add components to distilled/deion-
ized water and bring volume to 1.0L. Mix thoroughly. Store at 5°C.
Suspension is good for 6 months to 1 year.
Solution V:
Composition per 100.0mL:
Hemin ...........................................................................................0.2g
NH 4 OH, concentrated solution............................................ 2–3 drops
Preparation of Solution V: Add hemin to 10.0mL of distilled/de-
ionized water. Dissolve by adding a few drops of concentrated
NH 4 OH. Mix thoroughly. Bring volume to 100.0mL with distilled/de-
ionized water. Store at 5°C.
Solution VI:
Composition per 100.0mL:
DL -Thioctic acid.......................................................................10.0mg
Ethanol (95% solution)............................................................10.0mL
Preparation of Solution VI: Add thioctic acid to 10.0mL of 95%
ethanol. Mix thoroughly. Store at −20°C.
Solution VII:
Composition per 10.0mL:
Coenzyme A ............................................................................10.0mg
Na 2 S·5H 2 O (0.05% solution in freshly boiled,
distilled/deionized H 2 O).................................................... 2 drops
Preparation of Solution VII: Add coenzyme A to distilled/deion-
ized water and bring volume to 10.0mL. Mix thoroughly. Add 2 drops
of 0.05% Na 2 S·5H 2 O solution. Store at −20°C.
Preparation of Casein Hydrolysate-Vitamin-Glucose Base:
Add components to distilled/deionized water and bring volume to
450.0mL. Mix thoroughly. Adjust pH to 6.5 with 20% KOH solu-
tion. Bring volume to 500.0mL with distilled/deionized water. Mix
thoroughly. Filter sterilize.
Agar Starch Base:
Composition per 500.0mL:
Agar............................................................................................12.5g
Potato starch, insoluble.................................................................2.0g
Solution VIII............................................................................10.0mL
Solution VIII:
Composition per 100.0mL:
Oleic acid......................................................................................0.1g
Preparation of Solution VIII: Add oleic acid to distilled/deion-
ized water and bring volume to 50.0mL. Mix thoroughly. Adjust pH to
7.0 with NaOH. Bring volume to 100.0mL with distilled/deionized wa-
ter.
Preparation of Agar Starch Base: Add potato starch to distilled/
deionized water and bring volume to 50.0mL. Mix thoroughly. Pour
into 440.0mL of boiling, distilled/deionized water. Add 10.0mL of so-
lution VIII and the agar. Mix thoroughly. Autoclave for 20 min at 15
psi pressure–121°C.
Preparation of Medium: Aseptically combine 500.0mL of filter-
sterilized casein hydrolysate-vitamin-glucose base with 500.0mL of
hot agar starch base. Mix thoroughly. Heat nearly to boiling while stir-
ring constantly. Aseptically distribute into sterile tubes. Allow tubes to
cool in a slanted position.
Use: For the cultivation and maintenance of Blastomyces dermatitidis
and Histoplasma capsulatum in the yeast phase.