Isolation and fractionation of total nucleic acids from tissues and cells.
A simple and efficient method was devised to permit the isolation of total nucleic acids (poly(A+) and poly(A-) RNAs and DNA) from cells and tissues (e.g. testis) enriched in DNA. Chaotropic reagents (guanidine thiocyanate and LiBr) were utilized to inactivate nucleases rapidly, minimize the viscosity of the homogenization solution and to precipitate RNA selectively (LiBr). Both total and poly(A+)-enriched mRNAs were recovered in a biologically active form as demonstrated by their ability to programme an in vitro translation reaction. High molecular weight DNA (greater than 22 kilobases) was recovered from the LiBr-soluble supernatants by selective ethanol precipitation, subsequently purified and was in a form suitable for further biochemical analysis.
D000818:Animals; D001965:Bromides; D002417:Cattle; D002478:Cells, Cultured; D004247:DNA / Q000302:isolation & purification*; D019791:Guanidine; D006146:Guanidines; D008094:Lithium; D018020:Lithium Compounds; D008099:Liver / Q000032:analysis; D008297:Male; D014176:Protein Biosynthesis; D012333:RNA, Messenger / Q000302:isolation & purification*; D013737:Testis / Q000032:analysis
D001965:Bromides; D006146:Guanidines; D018020:Lithium Compounds; D012333:RNA, Messenger; C040949:lithium bromide; D004247:DNA; D008094:Lithium; D019791:Guanidine