Simultaneous extraction from clinical biopsies of high-molecular-weight DNA and RNA: comparative characterization by biotinylated and 32P-labeled probes on Southern and Northern blots.
A method for efficient simultaneous extraction of high-molecular-weight DNA and RNA from solid mammalian tissues including clinical biopsies is described. It is based on the disruption and subsequent melting of deep frozen tissue in the presence of frozen phenol and nucleic acid extraction buffer; this allows for simultaneous disruption of tissue and inactivation of nucleases. The yield is about 0.7-5.8 mg of DNA and 0.5-8.1 mg of total RNA/g of tissue depending upon the tissue type; this is higher than the yield of other methods tested. Analysis of total RNA by denaturing gel electrophoresis, and of DNA and poly(A)+ RNA by Southern and Northern blot hybridization using 32P and biotinylated probes, indicated that c-Ha-ras gene and its transcripts were undegraded. Biotinylated and 32P probes had approximately the same sensitivity in detecting nucleic acids on Southern and Northern blots. This extraction procedure is simple and, when used with biotinylated probes, is rapid, inexpensive, and nonhazardous. The methodology can be modified for use with other clinical samples and cells grown in culture.
D000818:Animals; D001710:Biotin; D001943:Breast Neoplasms / Q000032:analysis; D004247:DNA / Q000302:isolation & purification*; D004587:Electrophoresis, Agar Gel; D006801:Humans; D008099:Liver / Q000032:analysis; D008970:Molecular Weight; D009693:Nucleic Acid Hybridization; D010761:Phosphorus Radioisotopes; D012313:RNA / Q000302:isolation & purification*; D051381:Rats
D010761:Phosphorus Radioisotopes; D012313:RNA; D001710:Biotin; D004247:DNA