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Mapping Adenines, Guanines, And Pyrimidines In Rna, Helen Donis-Keller, Allan Maxam, Walter Gilbert
Mapping Adenines, Guanines, And Pyrimidines In Rna, Helen Donis-Keller, Allan Maxam, Walter Gilbert
Helen Donis-Keller
The positions of adenines, guanines, and pyrimidines can be determined by partial nuclease digestion of a terminally labeled RNA molecule. In urea, at elevated temperatures, RNase T1 generates a pattern reflecting cleavage at guanines while RNase U2 cleaves only at adenine. A limited alkaline hydrolysis provides a continuum of fragments derived from breaks at every phosphodiester bond. The reaction products are electrophoretically fractionated by size in adjacent lanes of a polyacrylamide gel. An autoradiograph of the gel displays the sequence up to 100 nucleotides from the end of the molecule, although uracil cannot as yet be distinguished from cytosine. These …
Separation Of Quantitation Of Intracellular Forms Of Poliovirus Rna By Agarose Gel Electrophoresis, Martinez Hewlett, Shmuel Rozenblatt, Victor Ambros, David Baltimore
Separation Of Quantitation Of Intracellular Forms Of Poliovirus Rna By Agarose Gel Electrophoresis, Martinez Hewlett, Shmuel Rozenblatt, Victor Ambros, David Baltimore
Victor R. Ambros
Intracellular poliovirus-specific RNA species can be measured directly by electrophoresis of total cytoplasmic nucleic acids through 1% agarose gels, resulting in the separation of single- and double-stranded forms of poliovirus RNA from each other and from HeLa cell 28S ribosomal RNA. Single-stranded RNA molecules differing by only 15% in length are resolved in this gel system. RNA species can be visualized as fluorescen bands appearing after staining of the gels with ethidium bromide and observation under ultraviolet illumination. The total amount of RNA can be determined by densitometric quantitation of the fluorescent response. In this way, the amount of poliovirus-specific …