Blood Agar, Diphasic

[所属分类:培养基配方] [发布时间:2019-12-26] [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

Blood Agar, Diphasic

培养基配方
Composition per 800.0mL:
Lean beef, desiccated.......................................25.0g
Agar .......................................................10.0g
Neopeptone..................................................10.0g
NaCl.........................................................2.5g
Locke solution............................................200.0mL
Rabbit blood, defibrinated ...............................100.0mL
pH 7.2–7.4 at 25°C
Locke Solution:
Composition per liter:
NaCl........................................................8.0g
Glucose ....................................................2.5g
KH2PO4 .....................................................0.3g
KCl.........................................................0.2g
CaCl2 ·2H2O................................................0.2g
Preparation of Locke Solution: Add components to distilled/de-
ionized water and bring volume to 1.0L. Mix thoroughly. Filter steril-
ize.
Preparation of Medium: Add beef to 500.0mL of distilled/deion-
ized water. Let stand for 60 min. Gently heat and bring to 80°C for 5
min. Filter through Whatman #1 filter paper. To filtrate, add remaining
components, except Locke solution and rabbit blood. Mix thoroughly.
Adjust pH to 7.2–7.4 with NaOH. Autoclave for 20 min at 15 psi pres-
sure–121°C. Cool to 45°–50°C. Aseptically add sterile rabbit blood.
Mix thoroughly. Aseptically distribute into sterile tubes in 5.0mL vol-
umes. Allow tubes to cool in a slanted position. Immediately prior to
inoculation, overlay agar in each tube with 2.0mL of sterile Locke so-
lution.
Use: For the cultivation of Trypanosoma species and Leishmania spe-
cies.