Published online 15 January 2004
Nucleic Acids Research, 2004, Vol. 32, No. 1 e14
© 2004 Oxford University Press
Superior 5' homogeneity of RNA from ATP-initiated transcription under the T7 
2.5 promoter
Department of Chemistry and Biochemistry, University of Southern Mississippi, Hattiesburg, MS 39406-5043, USA
*To whom correspondence should be addressed. Tel: +1 601 266 4371; Fax: +1 601 266 6075; Email: faqing.h.huang{at}usm.edu
The authors wish it to be known that, in their opinion, the first two authors should be regarded as joint First Authors
Transcription from the commonly used GTP- initiating T7 class III promoter 
6.5 frequently produces heterogeneous RNA at both 3' and 5' ends. We demonstrate here that RNA transcripts from the T7 class II promoter 2.5 have superior 5' homogeneity over those from the 6.5 promoter, with comparable total RNA yields. The overall homogeneity of RNA transcripts is improved to different degrees depending on RNA sequences, although transcription under 2.5 does not affect the 3' heterogeneity of RNA. In combination with 3' RNA trimming by DNAzymes or ribozymes, this ATP- initiated transcription system based on the T7 2.5 promoter can provide excellent quality of RNA for applications requiring a high degree of RNA size homogeneity.
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