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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 {phi}2.5 promoter

Tricia M. Coleman, Guocan Wang and Faqing Huang*

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 {phi}6.5 frequently produces heterogeneous RNA at both 3' and 5' ends. We demonstrate here that RNA transcripts from the T7 class II promoter {phi}2.5 have superior 5' homogeneity over those from the {phi}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 {phi}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 {phi}2.5 promoter can provide excellent quality of RNA for applications requiring a high degree of RNA size homogeneity.


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