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

*この研究の対応する著者

研究成果: Article査読

12 被引用数 (Scopus)

抄録

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.

本文言語English
ページ(範囲)1042-1049
ページ数8
ジャーナルBioTechniques
31
5
DOI
出版ステータスPublished - 2001
外部発表はい

ASJC Scopus subject areas

  • バイオテクノロジー
  • 生化学、遺伝学、分子生物学(全般)

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