An improved method for isolating RNA from coccidian oocysts.
A method, based on the disruption of eimerian oocysts in a French pressure cell in the presence of guanidine isothiocyanate, has been developed to isolate large quantities of high quality total RNA efficiently. This procedure results in a 12.5-fold greater number of oocysts broken, and a 22-fold greater yield of total RNA than from disruption by conventional grinding in liquid N2. In addition, the RNA isolated by the French pressure cell method was of equal or superior quality when compared to RNA isolated by grinding. This procedure provides a significant improvement in RNA extraction from eimerian oocysts and will greatly facilitate the study of gene expression in this important group of parasites.
Abrahamsen
M S
MS
Montana State University, Bozeman 59717.
Clark
T G
TG
White
M W
MW
D000818:Animals; D015152:Blotting, Northern; D003080:Cold Temperature; D004539:Eimeria / Q000235:genetics*; D006146:Guanidines; D017879:Isothiocyanates; D009584:Nitrogen; D011312:Pressure; D016053:RNA, Protozoan / Q000302:isolation & purification*
D006146:Guanidines; D017879:Isothiocyanates; D016053:RNA, Protozoan; C054435:guanidine isothiocyanate; D009584:Nitrogen