Source
Department of Biology, Indiana University, Bloomington 47405, USA. ckao@bio.indiana.edu
Abstract
DNA templates modified with C2'-methoxyls at the last two nucleotides of the 5' termini dramatically reduced nontemplated nucleotide addition by the T7 RNA polymerase from both single- and double-stranded DNA templates. This strategy was used to generate several different transcripts. Two of the transcripts were demonstrated by nuclear magnetic resonance spectroscopy to be unaffected in their sequence. Transcripts produced from the modified templates can be purified with greater ease and should be useful in a number of applications.