Improved simple chromatographic method for the fractionation of major classes of mammalian nucleic acids.
CL-Sepharose 4B column chromatography has been used for the separation of four major classes of mammalian nucleic acids in a single chromatographic run. Gel filtration at 2.5 M NaCl separated DNAs (containing RNA hybrids) from tRNAs. The 18 S RNA (containing 3-5 wt% of small 5 S RNA and RNA degradative products) was eluted at 0.7 M NaCl, and 28 S RNA (containing hnRNAs) was eluted at 0.1 M NaCl. Poly(A)+ mRNAs were detected in both 18 and 28 S RNA fractions. The present procedure is suitable for both analytical and preparative work and may serve as an initial step for the further isolation of ultrapure nucleic acid preparations.
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Popović
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Leskovac
D000818:Animals; D002286:Carcinoma, Ehrlich Tumor / Q000032:analysis; D002847:Chromatography, Agarose / Q000379:methods*; D002850:Chromatography, Gel / Q000379:methods*; D004247:DNA / Q000302:isolation & purification*; D004273:DNA, Neoplasm / Q000032:analysis; D007668:Kidney / Q000032:analysis; D008099:Liver / Q000032:analysis; D051379:Mice; D012313:RNA / Q000302:isolation & purification*; D012334:RNA, Neoplasm / Q000032:analysis; D012335:RNA, Ribosomal / Q000302:isolation & purification; D012343:RNA, Transfer / Q000302:isolation & purification; D051381:Rats; D012685:Sepharose / Q000031:analogs & derivatives; D013056:Spectrophotometry, Ultraviolet
D004273:DNA, Neoplasm; D012334:RNA, Neoplasm; D012335:RNA, Ribosomal; C035771:Sepharose CL 4B; D012313:RNA; D004247:DNA; D012685:Sepharose; D012343:RNA, Transfer