| US 7,585,631 B2 | ||
| Method for obtaining purified RNA from viable oocysts | ||
| Melissa M. Cunningham, Sliema (Malta); Paul D. Stull, Wilmington, Del. (US); and William G. Weisburg, San Diego, Calif. (US) | ||
| Assigned to Gen-Probe Incorporated, San Diego, Calif. (US) | ||
| Filed on Jul. 25, 2006, as Appl. No. 11/459,885. | ||
| Application 11/459885 is a continuation of application No. 09/945586, filed on Sep. 11, 2001, granted, now 7,081,527, filed on Jul. 25, 2006. | ||
| Claims priority of provisional application 60/232028, filed on Sep. 12, 2000. | ||
| Prior Publication US 2007/0020661 A1, Jan. 25, 2007 | ||
| Int. Cl. C12Q 1/68 (2006.01); C07H 21/00 (2006.01); C07H 21/02 (2006.01); C07H 21/04 (2006.01) | ||
| U.S. Cl. 435—6 [536/22.1; 536/23.1; 536/24.3] | 14 Claims |
| 1. A method for obtaining purified RNA from viable oocysts, the method comprising the steps of:
a) centrifuging a fluid sample containing oocysts at a speed and for a period of time sufficient to concentrate the oocysts
within a vessel containing the fluid sample;
b) removing a supernatant formed in the vessel during the centrifuging step;
c) resuspending the concentrated oocysts in a buffered solution;
d) agitating the buffered solution in the presence of zirconia/silica particles at a rate and for a period of time sufficient
to lyse the oocysts and release RNA therefrom;
e) immobilizing the released RNA on an RNA-binding filter;
f) purifying the released RNA by washing the filter one or more times with a buffered solution to remove oocyst components
other than the released RNA; and
g) removing the purified RNA from the filter.
|