Simultaneous isolation of total cellular RNA and DNA from tissue culture cells using phenol and lithium chloride.
A rapid procedure for the isolation of intact total cellular RNA from cultured cells is described. This method combines the simultaneous disruption of cells and extraction of nucleic acids in a single step with the use of phenol and a buffer containing 100 mM LiCl. Total cellular RNA can be isolated in approximately 2 hours. The yield and quality of the RNA is comparable to the more widely employed methods requiring extensive preparatory steps such as extraction using guanidinium thiocyanate and subsequent CsCl gradient centrifugation. The RNA isolated using our procedure contains transcripts up to 10 kb in length and is suitable for Northern analysis. This procedure also yields high-molecular-weight DNA, which is a suitable substrate for restriction endonucleases.
Raha
S
S
Department of Biochemistry, Erindale College, University of Toronto, Mississauga, Ontario, Canada.
Merante
F
F
Proteau
G
G
Reed
J K
JK
D000310:Adrenal Gland Neoplasms / Q000737:chemistry; D000818:Animals; D015152:Blotting, Northern; D015139:Blotting, Southern; D002499:Centrifugation, Density Gradient; D002586:Cesium; D002712:Chlorides; D046508:Culture Techniques; D004247:DNA / Q000302:isolation & purification*; D004262:DNA Restriction Enzymes; D006146:Guanidines; D008094:Lithium; D018021:Lithium Chloride; D009132:Muscles / Q000737:chemistry; D010636:Phenols; D010673:Pheochromocytoma / Q000737:chemistry; D012313:RNA / Q000302:isolation & purification*; D051381:Rats; D013861:Thiocyanates; D014407:Tumor Cells, Cultured / Q000737:chemistry
D002712:Chlorides; D006146:Guanidines; D010636:Phenols; D013861:Thiocyanates; D002586:Cesium; C054436:guanidine thiocyanate; D012313:RNA; D004247:DNA; D008094:Lithium; D004262:DNA Restriction Enzymes; D018021:Lithium Chloride; C028019:cesium chloride