Diphasic Medium for Amoeba (Charcoal Agar Slants)

[所属分类:培养基配方] [发布时间:2020-6-22] [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

Diphasic Medium for Amoeba
(Charcoal Agar Slants)

培养基配方
Composition per liter:
Agar slants................................................................1.0L
Buffered saline overlay....................................................1.0L
pH 7.4 ± 0.2 at 25°C
Agar Slants:
Composition per liter:
Agar........................................................................10.0g
Charcoal, activated ........................................................10.0g
Pancreatic digest of casein..................................................5.0g
KH2PO4 ......................................................................4.0g
Na2HPO4 .....................................................................3.0g
Asparagine...................................................................2.0g
Sodium citrate...............................................................1.0g
Ferric ammonium citrate......................................................0.1g
MgSO4 ·7H2O.................................................................0.1g
Cholesterol solution.......................................................25.0mL
Glycerol ..................................................................10.0mL
Cholesterol Solution:
Composition per 25.0mL:
Cholesterol.................................................................0.25g
Acetone.....................................................................25.0mL
Preparation of Cholesterol Solution: Add cholesterol to 25.0mL
of acetone. Mix thoroughly.
Preparation of Agar Slants:  Add components, except agar, char-
coal, and cholesterol solution, to distilled/deionized water and bring
volume to 1.0L. Mix thoroughly. Gently heat to dissolve. Do not boil.
Add agar, charcoal, and cholesterol solution. Mix thoroughly. Gently
heat and bring to boiling. Distribute into tubes in 3.0mL volumes. Au-
toclave for 15 min at 15 psi pressure–121°C. Resuspend charcoal. Al-
low tubes to cool in a slanted position with short butts or no butts.
Buffered Saline Overlay:
Composition per liter:
NaCl............................................................................5.0g
Solution B...................................................................810.0mL
Solution A...................................................................190.0mL
Solution A:
Composition per liter:
KH2PO4 , anhydrous............................................................9.07g
Preparation of Solution A: Add KH2PO4 to distilled/deionized
water and bring volume to 1.0L. Mix thoroughly.
Solution B:
Composition per liter:
Na2HPO4 , anhydrous...........................................................9.46g
Preparation of Solution B: Add Na2HPO4 to distilled/deionized
water and bring volume to 1.0L. Mix thoroughly.
Preparation of Buffered Saline Overlay: Combine 810.0mL of
solution A and 190.0mL of solution B. Add the NaCl. Mix thoroughly.
Autoclave for 15 min at 15 psi pressure–121°C. Cool to 25°C. Store at
4°C.
Preparation of Medium: To each agar slant, aseptically add 3.0mL
of sterile, buffered saline overlay.
Use: For the cultivation and maintenance of Amoebae species.