Sequence Analysis of Nonradioactive RNA Fragments by Periodate-phosphatase Digestion and Chemical Tritium Labeling: Characterization of Large Oligonucleotides and Oligonucleotides Containing Modified Nucleosides

Overview

Journal Nucleic Acids Res

Publisher Oxford University Press

Specialty Biochemistry

Date 1974 Sep 1

PMID 4375807

Citations 11

Authors

K Randerath

E RANDERATH

L S Chia

R C Gupta

M Sivarajan

Affiliations

Soon will be listed here.

Abstract

A tritium derivative method for sequence analysis of polyribonucleotides is detailed, which is based on borotritide reduction of oligonucleotide dialdehydes generated by treatment of polyribonucleotides with alkaline phosphatase and excess periodate at pH 8 (borate buffer; no primary amine present in the reaction mixture). While neither phosphatase nor periodate possess any intrinsic exonuclease activity their combination mimics an RNA-specific exonuclease ("pseudo-exonuclease"). Procedures are described for separation and characterization of tritiumlabeled oligonucleotide derivatives. The sequence is deduced by identification of labeled 3'-termini following separation of the reduced nucleotide intermediates according to chain length. The sensitivity of the method is indicated by the fact that as little as 0.01 O.D.260 unit of a nonradioactive decanucleotide is sufficient for sequence determination.

Citing Articles

Sequence studies of nonradioactive Mycoplasma tRNA Phe with the aid of polynucleotide phosphorylase and polynucleotide kinase.

Szeto K, Soll D Nucleic Acids Res. 1974; 1(12):1733-8.

PMID: 4449733 PMC: 343452. DOI: 10.1093/nar/1.12.1733.

A new method for sequence analysis of oligodeoxyribonucleotides.

Black D, GILHAM P Nucleic Acids Res. 1985; 13(7):2433-42.

PMID: 2987853 PMC: 341165. DOI: 10.1093/nar/13.7.2433.

An improved separation procedure for nucleoside monophosphates on polyethyleneimine-(PEI-)cellulose thin layers.

Gupta R, RANDERATH E, Randerath K Nucleic Acids Res. 1976; 3(11):2915-21.

PMID: 1005109 PMC: 343141. DOI: 10.1093/nar/3.11.2915.

Use of specific endonuclease cleavage in RNA sequencing-an enzymic method for distinguishing between cytidine and uridine residues.

Gupta R, Randerath K Nucleic Acids Res. 1977; 4(10):3441-54.

PMID: 928065 PMC: 342663. DOI: 10.1093/nar/4.10.3441.

A double-labeling procedure for sequence analysis of picomole amounts of nonradioactive RNA fragments.

Gupta R, RANDERATH E, Randerath K Nucleic Acids Res. 1976; 3(11):2895-914.

PMID: 826884 PMC: 343140. DOI: 10.1093/nar/3.11.2895.

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