Open AccessPreparation of Nascent RNA Strands of the Bacteriophage M12
Author(s) -
Francke Bertold,
Ward Richard,
Konings Rund N. H.,
Hofschneider Peter Hans
Publication year - 1970
Publication title -
european journal of biochemistry
Language(s) - English
Resource type - Journals
eISSN - 1432-1033
pISSN - 0014-2956
DOI - 10.1111/j.1432-1033.1970.tb01038.x
Subject(s) - rna , bacteriophage , centrifugation , sucrose gradient , ultracentrifuge , nucleotide , chemistry , sedimentation coefficient , sucrose , dna , sedimentation , biology , microbiology and biotechnology , biochemistry , biophysics , escherichia coli , gene , membrane , enzyme , paleontology , sediment
Nascent M12 RNA has been isolated in large quantities from Replicative Intermediate and denatured by a short heat treatment. After sucrose gradient centrifugation of the denatured Replicative Intermediate in a zonal rotor, the nascent RNA strands were divided into six fractions with increasing sedimentation values. Each fraction was further purified by sedimentation conventional sucrose gradients. In this manner, a set of M12 nascent RNA strands with fairly uniform lengths, ranging from about 15 nucleotides to almost 90% of the complete phage RNA genome, have been obtained. The molecules in these fractions, all containing the same 5′‐end as complete phage RNA but an increasing portion of the phage genome, provide suitable tools for studies of the structure and function of phage RNA.