L Diphasic Blood Agar Medium (ATCC Medium 1011)

[所属分类:培养基配方] [发布时间:2021-4-25] [发布人:李佳颐] [阅读次数:] [返回]

L Diphasic Blood Agar Medium
(ATCC Medium 1011)

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

Composition per 1150.0mL:

Blood agar, diphasic base medium ........................................700.0mL
Rabbit blood, defibrinated .....................................................300.0mL
Locke’s solution.....................................................................150.0mL
pH 7.2–7.4 at 25°C
Blood Agar, Diphasic Base Medium:
Composition per 750.0mL:
Beef.............................................................................................25.0g
Agar ............................................................................................10.0g
Neopeptone.................................................................................10.0g
NaCl..............................................................................................2.5g
Preparation of Blood Agar, Diphasic Base Medium: Trim
beef to remove fat. Add 25.0g of lean beef to 250.0mL of distilled/de-
ionized water. Gently heat and bring to boiling. Boil for 2–3 min. Filter
through Whatman #2 filter paper. Add agar, neopeptone, and NaCl to
filtrate. Bring volume to 750.0mL with distilled/deionized water. Mix
thoroughly. Adjust pH to 7.2–7.4. Gently heat and bring to boiling. Au-
toclave for 15 min at 15 psi pressure–121°C. Cool to 50°–55°C.
Locke’s Solution:
Composition per liter:
NaCl..............................................................................................8.0g
Glucose .........................................................................................2.5g
KH 2 PO 4 ........................................................................................ 0.3g
CaCl 2 ............................................................................................ 0.2g
KCl................................................................................................0.2g
Preparation of Locke’s Solution:  Add components to distilled/
deionized water and bring volume to 1.0L. Mix thoroughly. Autoclave
for 15 min at 15 psi pressure–121°C. Cool to 50°–55°C.
Preparation of Medium: Aseptically combine 700.0mL of sterile
blood agar, diphasic base medium, with 300.0mL of sterile defibrinat-
ed rabbit blood warmed to 50°–55°C. Mix thoroughly. Aseptically dis-
tribute 5.0mL volumes into 16 × 125mm screw-capped test tubes.
Allow to cool in a slanted position. Overlay the agar in each tube with
3.0mL of sterile Locke’s solution.
Use:  For the cultivation and maintenance of Trypanosoma species.