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

• Published in: Journal of biochemistry (Tokyo) 1972-09, Vol.72 (3), p.561-570
• Main Authors: ABE, Sachiko, FUJISAWA, Takao, SATAKE, Mei, OGATA, Kikuo
• Format: Article
• Language: English
• ISSN: 0021-924X; 1756-2651
• DOI: 10.1093/oxfordjournals.jbchem.a129935

Rat liver nuclei were labeled in vivo with 14C-orotic acid for 30 min, and the effects of SDS,** temperature, pH and ionic strength during SDS-phenol extraction on the recovery, specific activity, and extents of contamination of the nuclear RNA with DNA and protein were investigated. The standard extraction medium contained 0.1% SDS, 0.14 M NaCl, and 0.025 M sodium acetate buffer (pH 5.1). 1. On increasing the SDS concentration from 0 to 1%, the recovery and specific activity of nuclear RNA increased, especially at 30°C. Extraction at 65°C was slightly better than at 30°C with the same concentrations of SDS. 2. Using single and serial extraction methods a rise of temperature from 30 to 75°C increased the specific activity of nuclear RNA by about 2 and 8 fold, respectively. 3. Increases in temperature and SDS concentration had similar effects on extraction of extranucleolar RNA. Nucleolar RNA was as effectively extracted with 0.1% SDS-phenol at 30°C as at 65°C. 4. NaCl decreased the specific activity of nuclear RNA and had little effect on its recovery at either 30 or 65°C. 5. Increase in the NaCl or SDS concentration increased contamination with protein and DNA at 30°C but not at 65°C. 6. Change in pH had little effect on any of these factors at either temperature.