A simple and efficient method for purifying and quantifying schizonts from Theileria parva-infected cells.
An improved method for the purification of Theileria parva schizonts from infected bovine cells is described. The technique is simpler and more rapid than previously described methods and gives rise to greater yields of schizonts with negligible contamination by host-cell components. In addition, a fluorescent staining technique was developed whereby live schizonts purified from infected cells can be enumerated and sorted using the flow cytometer. An assessment of the quality of schizonts prepared according to our method as a source of RNA for the construction of parasite cDNA libraries suggests that RNA derived from these preparations is free of host nucleic acids.
International Laboratory for Research on Animal Diseases, Nairobi, Kenya.
B M
BM
Goddeeris
S
S
Dunlap
E A
EA
Innes
D J
DJ
McKeever
D000818:Animals; D015152:Blotting, Northern; D002417:Cattle; D002460:Cell Line; D005434:Flow Cytometry; D016053:RNA, Protozoan / Q000032:analysis* / Q000302:isolation & purification; D016797:Theileria parva / Q000235:genetics / Q000254:growth & development / Q000302:isolation & purification*
D016053:RNA, Protozoan