EXTRACTION OF RNA FROM RAT UTERUS

Methods for the extraction of RNA from the immature rat uterus have been critically evaluated. Primary emphasis was placed upon minimizing degradation of RNA. The procedure of choice consists of extraction with phenol and sodium dodecyl sulfate at 50° in the presence of bentonite, followed by deoxyribonuclease treatment of the extracted material. The product contains an average of 55% of the total RNA and >26% of the rapidly-labeled RNA (20-min pulse of [5-3H]uridine) of the uterus. (The recoveries are based on values obtained by hot HClO4 extraction of RNA.) Almost all of the rapidly-labeled RNA in the product is of high molecular weight, with sedimentation coefficients in the range 30-50 S on sucrose gradients, which suggests that degradation of the material has been minimized. Estradiol given intraperitoneally to immature rats 2, 4, or 12 h before sacrifice did not affect the pattern of distribution of this high-molecular-weight, rapidly-labeled RNA (10-, 20-, or 40-min pulse of [5-3H]uridine) on sucrose gradients. Estradiol given intraperitoneally 20 min before sacrifice to rats ovariectomized at 28 days and used for experimentation 18-33 days after ovariectomy also did not affect the pattern of distribution of rapidly-labeled RNA (10-min pulse of [5-3H]uridine) on sucrose gradients. © 1969.