Download RNA Processing Part A: General Methods by Melvin I. Simon, James E. Dahlberg, John N. Abelson PDF

By Melvin I. Simon, James E. Dahlberg, John N. Abelson

Many equipment were constructed or tailored for the examine of RNA processing, which performs a key function within the expression of genetic details. Volumes a hundred and eighty and 181 of Methods in Enzymology, RNA Processing, elements A and B, symbolize the 1st finished collation of protocols used for the training of substrates, for the education of extracts and enzymes, and for the research of goods in RNA processing. a minimum of 1/2 the chapters current concepts which are without delay appropriate to different components of nucleic acid biochemistry and telephone biology

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L. Birnstiel, J. Mol. Biol. 118, 61 (1978). 7 R. De Wachter and W. Fiers, Anal. Biochem. 49, 184 (1972). METHODS IN ENZYMOLOGY. VOL. 180 Copyright © 1989 by Academic Press, inc. All rights of reproduction in any form reserved. [2] GEL ELECTROPHORESIS OF R N A 15 FIG. I. Two-dimensional gel separation of [32p]tRNAs and their precursors. An E. coli temperature-sensitive mutant (TS709) defective in RNase P at high temperature was labeled by 32PO~- at a permissive (a) or a restrictive (b) temperature.

Maxwell, and W. E. Hahn, Anal. Biochem. 72, 413 (1976). 36 PREPARATION OF SUBSTRATES [3] column, ethanol precipitated, and resuspended in Tris buffer. 2% (w/v) SDS. 5 M NaCI, and reloaded onto the oligo(dT) column. RNA concentrations less than 2 mg/ml minimize aggregation. 71 The binding properties of poly(A)-containing RNAs and ribonucleoprotein particles (RNPs) have been investigated as a function of monovalent and divalent ion concentration. 77 Poly(A) RNPs have similar binding properties to poly(A) RNAs in buffers containing 40 mM NaCI and 5 mM MgCI2.

Baguley, and R. K. Ralph, this seris, Vol. 12, Part A, p. 660. 28 PREPARATION OF SUBSTRATES [3] Benzoylated DEAE-Cellulose Esterification of the hydroxyl groups of cellulose with benzoic acid or with a combination of benzoic and naphthoic acid substantially improves the resolution of DEAE-cellulose by increasing the hydrophobic characteristics of the column. 17,18 Benzoylated DEAE-cellulose (BD-cellulose) and benzoylated, naphthoylated DEAE-cellulose (BND-cellulose) have similar properties to DEAE-cellulose, except that the binding interactions are generally stronger and that fractionation is more dependent on the secondary structure of the nucleic a c i d .

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