A simple, efficient method for the separate isolation of RNA and DNA from the same cells.
A method for the isolation of total cytoplasmic RNA and high molecular weight DNA from the same cells is described. Cells are gently lysed with NP40 in the presence of vanadyl ribonucleoside complex and the nuclei pelleted by centrifugation. RNA is purified by phenol/CHCl3 extraction of the lysate supernatant followed by ethanol precipitation. Protein is removed from high molecular weight DNA by salt-precipitation after nuclei are digested with proteinase K in the presence of sodium dodecyl sulfate. High yields of clean, intact RNA and DNA are obtained. A major advantage of the method is that it can be scaled down to quantitatively extract RNA and DNA from as little as 1000 cells.
Nicolaides
N C
NC
Department of Anatomy, University of Pennsylvania, School of Medicine, Philadelphia 19104.
Stoeckert
C J
CJ
D000818:Animals; D002460:Cell Line; D003593:Cytoplasm / Q000032:analysis; D004247:DNA / Q000302:isolation & purification*; D005347:Fibroblasts; D006801:Humans; D008722:Methods; D051379:Mice; D008970:Molecular Weight; D012313:RNA / Q000302:isolation & purification*; D012965:Sodium Chloride; D014407:Tumor Cells, Cultured
D012965:Sodium Chloride; D012313:RNA; D004247:DNA