Superior 5' Homogeneity of RNA from ATP-Initiated Transcription Under the T7 Φ2.5 Promoter

Authors

Tricia M. Coleman, University of Southern MississippiFollow
Guocan Wang, University of Southern MississippiFollow
Faqing Huang, University of Southern MississippiFollow

Document Type

Article

Publication Date

1-1-2004

Department

Chemistry and Biochemistry

Abstract

Transcription from the commonly used GTP- initiating T7 class III promoter phi6.5 frequently produces heterogeneous RNA at both 3' and 5' ends. We demonstrate here that RNA transcripts from the T7 class II promoter phi2.5 have superior 5' homogeneity over those from the phi6.5 promoter, with comparable total RNA yields. The overall homogeneity of RNA transcripts is improved to different degrees depending on RNA sequences, although transcription under phi2.5 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 phi2.5 promoter can provide excellent quality of RNA for applications requiring a high degree of RNA size homogeneity.

Comments

Originally published in Nucleic Acids Research, 2004, Vol. 32, No. 1

Publication Title

Nucleic Acids Research

Volume

32

Issue

1

Recommended Citation

Coleman, T. M., Wang, G., Huang, F. (2004). Superior 5' Homogeneity of RNA from ATP-Initiated Transcription Under the T7 Φ2.5 Promoter. Nucleic Acids Research, 32(1).
Available at: https://aquila.usm.edu/fac_pubs/9123