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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

Published online by Cambridge University Press:  01 September 1999

C. KAO ,
M. ZHENG  and
S. RÜDISSER
C. KAO
Affiliation:
Department of Biology, Indiana University, Bloomington, Indiana 47405, USA
M. ZHENG
Affiliation:
Department of Chemistry, University of California at Berkeley, and Physical Bioscience Division, Berkeley Lawrence National Laboratory, Berkeley, California, USA
S. RÜDISSER
Affiliation:
Department of Chemistry, University of California at Berkeley, and Physical Bioscience Division, Berkeley Lawrence National Laboratory, Berkeley, California, USA
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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.

Keywords

nontemplated nucleotide additionnucleotide analogT7 RNA polymerasetranscription
Type
METHODS
Information
RNA , Volume 5 , Issue 9 , September 1999 , pp. 1268 - 1272
Copyright
© 1999 RNA Society

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