Studies on SDS-Phenol Methods for Extraction of Rat Liver Nuclear RNA

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Abstract
The effects of various conditions for extraction of liver with SDS-phenol on the pattern of nuclear RNA on acrylamide gel electrophoresis were examined. 1. On extraction with hot SDS-phenol, 29.5S RNA (nucleolar) disappeared and the amounts of 28S RNA and 18S RNA's decreased with concomitant appearance of degraded RNA distributed between 28 and 10S. Heat-treatment of nuclear RNA extracted with cold (30°C) SDS-phenol, resulted in a similar pattern of degradation. 2. Polydispersed RNA, which was distributed between the origin and the region of about 10S and which is thought to be DNA-like RNA, was extracted better with hot SDS-phenol than with cold SDS-phenol. At concentrations of SDS above 0.3%, however, the polydispersed RNA was extracted well, even, with cold SDS-phenol. 3. With cold 0.1% SDS-phenol, nucleolar 32S and 29.5S RNA's, as well as the polydispersed RNA, were extracted best at pH 6.5 and second best at pH 8. NaCl tended to inhibit the extraction of polydispersed RNA. 4. RNA extracted from the extranucleolar fraction with cold 1% SDS-phenol contained more polydispersed RNA than that extracted with cold 0.1% SDS-phenol. The effective extraction of this polydispersed RNA by high concentrations of SDS was confirmed by the “serial SDS-phenol extraction method.” Hot SDS-phenol was effective for extraction of polydispersed RNA from the extranucleolar fraction, but caused degradation of 28S and 18S RNA in nuclear RNA.

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