Diphasic Blood Agar Medium with 30% 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 30% Blood

培养基配方
Composition per 1450.0mL:
Blood agar, diphasic base medium ........................................700.0mL
Locke’s solution........................................................450.0mL
Rabbit blood, defibrinated ..............................................300.0mL
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 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 Blastocrithidia culicis,
Crithidia deanei, Crithidia flexonema, Crithidia luciliae, Crithidia mellifi-
cae, Endotrypanum species, Herpetomonas anglusteri, Herpetomonas
mariadeanei, Herpetomonas megaseliae, Herpetomonas muscarum, Her-
petomonas roitmani, Leishmania braziliensis, Leishmania donovani,
Leishmania peruviana, Leishmania tarentolae, Leptomonas collosoma,
Leptomonas costoris, Leptomonas lactosovorans, Leptomonas mirabilis,
Leptomonas pulexsimulantis, Leptomonas samueli, Leptomonas sey-
mouri, Trypanosoma avium, Trypanosoma bennetti, Trypanosoma cervi,
Trypanosoma chattoni, Trypanosoma conorrhini, Trypanosoma cruzi,
Trypanosoma cyclops, Trypanosoma fallisi, Trypanosoma lewisi, Try-
panosoma lucknowi, Trypanosoma mega, Trypanosoma musculi, Try-
panosoma neveulemairei, Trypanosoma ranarum, Trypanosoma rotato-
rium, and Trypanosoma tamiasi.