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nontemplated nucleotide addition

RNA (1)

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A simple and efficient method to reduce nontemplated nucleotide addition at the 3′ terminus of RNAs transcribed by T7 RNA polymerase
C. KAO, M. ZHENG, S. RÜDISSER
Journal:

RNA / Volume 5 / Issue 9 / September 1999

Published online by Cambridge University Press:

01 September 1999, pp. 1268-1272

Print publication:

September 1999
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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.