Removal of polyA tails from full-length cDNA libraries for high-efficiency sequencing

Y. Shibata, P. Carninci*, K. Sato, N. Hayatsu, T. Shiraki, Y. Ishii, T. Arakawa, A. Hara, N. Ohsato, M. Izawa, K. Aizawa, M. Itoh, K. Shibata, A. Shinagawa, J. Kawai, Y. Ota, S. Kikuchi, N. Kishimoto, M. Muramatsu, Y. Hayashizaki

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

12 Citations (Scopus)


We have developed a method to overcome sequencing problems caused by the presence of homopolymer stretches, such as polyA/T, in cDNA libraries. PolyA tails are shortened by cleaving before cDNA cloning with type IIS restriction enzymes, such as GsuI, placed next to the oligo-dT used to prime the polyA tails of mRNAs. We constructed four rice Cap-Trapper-selected, full-length normalized cDNA libraries, of which the average residual polyA tail was 4 bases or shorter in most of the clones analyzed. Because of the removal of homopolymeric stretches, libraries prepared with this method can be used for direct sequencing and transcriptional sequencing without the slippage observed for libraries prepared with currently available methods, thus improving sequencing accuracy, operations, and throughput.

Original languageEnglish
Pages (from-to)1042-1049
Number of pages8
Issue number5
Publication statusPublished - 2001
Externally publishedYes

ASJC Scopus subject areas

  • Biotechnology
  • Biochemistry, Genetics and Molecular Biology(all)


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