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