Diphasic Blood Agar Medium with 10% Blood

[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 Blood Agar Medium with 10% Blood

培养基配方
Composition per 1120.0mL:
Blood agar, diphasic base medium ...........................................630.0mL
Locke’s solution...........................................................420.0mL
Rabbit blood, defibrinated ..................................................70.0mL
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
KH2PO4 .........................................................................0.3g
CaCl2 ..........................................................................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 630.0mL of sterile
blood agar, diphasic base medium, with 70.0mL of sterile defibrinated
rabbit blood warmed to 50°–55°C. Mix thoroughly. Aseptically distrib-
ute 5.0mL volumes into16 × 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 of Leishmania braziliensis, Leishmania enri-
ettii, Leishmania tropica, Trypanosoma conorrhini, Trypanosoma
cruzi, and Trypanosoma rangeli.