Rapid and quantitative preparation of cytoplasmic RNA from small numbers of cells.
An extremely simple procedure for preparing cytoplasmic RNA from small numbers of cells is described. Cells are lysed with the detergent NP-40 and efficient extraction of protein from the postnuclear cytoplasmic lysate is ensured by denaturation with sodium dodecyl sulfate and urea. This procedure is suitable for preparing RNA from many cell types. All procedures have been scaled down to be performed in 1.5-ml microfuge tubes and thus RNA may be prepared from small numbers of cells. The procedure is extremely rapid and RNA is ready for Northern gel analysis in less than 30 min. Because so few steps are involved, RNA recovery is quantitative.
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Walter and Eliza Hall Institute of Medical Research, Royal Melbourne Hospital, Victoria, Australia.
D000818:Animals; D015152:Blotting, Northern; D002452:Cell Count; D002478:Cells, Cultured; D003593:Cytoplasm / Q000032:analysis; D017830:Octoxynol; D011092:Polyethylene Glycols; D012313:RNA / Q000302:isolation & purification*; D012967:Sodium Dodecyl Sulfate; D014508:Urea
D012967:Sodium Dodecyl Sulfate; D011092:Polyethylene Glycols; D012313:RNA; D014508:Urea; D017830:Octoxynol; C010615:Nonidet P-40