Open AccessISOLATION OF LARGE OLIGONUCLEOTIDE FRAGMENTS FROM THE ALANINE RNA
Author(s) -
Jean Apgar,
George A. Everett,
Robert W. Holley
Publication year - 1965
Publication title -
proceedings of the national academy of sciences
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 5.011
H-Index - 771
eISSN - 1091-6490
pISSN - 0027-8424
DOI - 10.1073/pnas.53.3.546
Subject(s) - induced pluripotent stem cell , myocyte , in vitro , microbiology and biotechnology , oligonucleotide , stem cell , computational biology , cell , biology , drug discovery , cardiac electrophysiology , neuroscience , electrophysiology , bioinformatics , biochemistry , embryonic stem cell , dna , gene
The discovery that very brief treatment of the alanine RNA with RNase T1 cleaves the RNA. specifically into two large fragments' suggested that this enzyme might give additional specific cleavages if digestion conditions were a little more vigorous. The isolation of several large oligonucleotides resulting from such specific cleavages is the subject of this communication. Yeast alanine RNA2 was digested with 225 units of RNase T1 (Sankyo, Ltd., Tokyo) per mg of RNA. in 0.2 N pH 7.5 Tris buffer for 1 hr at 0WC; and, after removal of the RNase Ti by phenol extraction, the digest was chromatographed on DEAE-cellulose in 7 M urea. The chromatographic pattern is shown in Figure 1. For comparison, a pattern obtained after complete digestion of the alanine RNA
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