Low molecular weight RNA species from chromatin.
Several methods of preparing low molecular weight RNA from chick embryo chromatin have been examined. Traditional methods for dissociating chromatin utilizing high concentrations of salt (greater than 2 M) followed by high-speed centrifugation resulted in very low yields of RNA. Increased yields of RNA were obtained by treating chromatin at lower salt concentration (0.2-0.5 M). By using low salt extraction and sodium dodecyl sulfate-phenol deproteinization, six to eight low molecular weight homogeneous RNA species were isolated from chick embryo chromatin and mouse myeloma chromatin. In the myeloma system, all these RNAs are metabolically stable. Each component is homogeneous as examined by gel electrophoresis and hybridizes with mouse DNA at a rate consistent with a single species. There are multiple gene copies for these RNA species in the mouse genome, varying from 100 to 2000 copies for the different species. One of these RNAs is identical with 5S rRNA. In addition, the redundancy of genes for 18S, 28S, and 5S rRNA and tRNA was determined. Approximately 300 copies for 18 and 28S rTRNA and 500 copies for 5S rRNA were found. tRNAs were on an average 110-fold redundant with about 55 different species measured.
D000818:Animals; D002460:Cell Line; D002642:Chick Embryo; D002843:Chromatin / Q000032:analysis*; D003840:Deoxycholic Acid; D004591:Electrophoresis, Polyacrylamide Gel; D005069:Evaluation Studies as Topic; D006146:Guanidines; D007700:Kinetics; D008722:Methods; D051379:Mice; D008970:Molecular Weight; D009691:Nucleic Acid Denaturation; D009693:Nucleic Acid Hybridization; D009994:Osmolar Concentration; D012313:RNA / Q000302:isolation & purification*; D012334:RNA, Neoplasm / Q000302:isolation & purification*; D012967:Sodium Dodecyl Sulfate; D013696:Temperature; D014461:Ultracentrifugation
D002843:Chromatin; D006146:Guanidines; D012334:RNA, Neoplasm; D003840:Deoxycholic Acid; D012967:Sodium Dodecyl Sulfate; D012313:RNA