PRAS-PYG with Tween™ 80

[所属分类:培养基配方] [发布时间:2021-7-13] [发布人:网站管理员2] [阅读次数:] [返回]

PRAS-PYG with Tween™ 80

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

Composition per 1054.25mL:

Glucose.......................................................................................10.0g
Yeast extract................................................................................10.0g
Peptone .........................................................................................5.0g
Pancreatic digest of casein............................................................5.0g
L -Cysteine·HCl·H 2 O.....................................................................0.5g
Salts solution............................................................................40.0mL
Hemin solution.........................................................................10.0mL
Resazurin 0.025%......................................................................4.0mL
Tween™ 80..............................................................................0.25mL
Vitamin K 1 solution...................................................................0.2mL
pH 7.0 ± 0.2 at 25°C
Salts Solution:
Composition per liter:
NaHCO 3 ......................................................................................10.0g
NaCl..............................................................................................2.0g
K 2 HPO 4 .........................................................................................1.0g
KH 2 PO 4 .........................................................................................1.0g
CaCl 2  (anhydrous).........................................................................0.2g
MgSO 4 ..........................................................................................0.2g
Preparation of Salts Solution: Dissolve CaCl 2 and MgSO 4 in
300.0mL of distilled water. Add 500.0mL of water, and add the remain-
ing salts while swirling slowly. Add 200.0mL of distilled water, mix,
and store at 4°C.
Hemin Solution:
Composition per 100.0mL:
Hemin.......................................................................................50.0mg
NaOH (1N solution)...................................................................1.0mL
Preparation of Hemin Solution: Dissolve hemin in 1.0mL of 1N
NaOH solution. Bring volume to 100.0mL with distilled/deionized wa-
ter. Mix thoroughly. Autoclave for 15 min at 15 psi pressure–121°C.
Vitamin K 1 Solution:
Composition per 30.15mL:
Ethanol (95% solution)............................................................30.0mL
Vitamin K 1 ...............................................................................0.15mL
Preparation of Vitamin K 1 Solution: Combine components. Mix
thoroughly. Store at 4°C in the dark. Discard solution after 1 month.
Preparation of Medium: Prepare and dispense medium under 97%
N 2 + 3% H 2 . Add components, except glucose, Tween™ 80, and  L -
cysteine·HCl·H 2 O, to distilled/deionized water and bring volume to 1.0L.
Mix thoroughly. Gently heat and bring to boiling. Continue boiling for 3
min. Cool to room temperature while sparging with 97% N 2 + 3% H 2 .
Add glucose, Tween™ 80, and  L -cysteine·HCl·H 2 O. Mix thoroughly.
Anaerobically distribute into sterile tubes or flasks. Autoclave for 15 min
at 15 psi pressure–121°C.
Use: For the cultivation of Clostridium puniceum.