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

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

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 pol...

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Veröffentlicht in:BioTechniques. - 1993. - 31(2001), 5 vom: 16. Nov., Seite 1042, 1044, 1048-9
1. Verfasser: Shibata, Y (VerfasserIn)
Weitere Verfasser: Carninci, P, Sato, K, Hayatsu, N, Shiraki, T, Ishii, Y, Arakawa, T, Hara, A, Ohsato, N, Izawa, M, Aizawa, K, Itoh, M, Shibata, K, Shinagawa, A, Kawai, J, Ota, Y, Kikuchi, S, Kishimoto, N, Muramatsu, M, Hayashizaki, Y
Format: Aufsatz
Sprache:English
Veröffentlicht: 2001
Zugriff auf das übergeordnete Werk:BioTechniques
Schlagworte:Journal Article Research Support, Non-U.S. Gov't DNA, Complementary Poly A 24937-83-5
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520 |a 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 
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650 4 |a Research Support, Non-U.S. Gov't 
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700 1 |a Carninci, P  |e verfasserin  |4 aut 
700 1 |a Sato, K  |e verfasserin  |4 aut 
700 1 |a Hayatsu, N  |e verfasserin  |4 aut 
700 1 |a Shiraki, T  |e verfasserin  |4 aut 
700 1 |a Ishii, Y  |e verfasserin  |4 aut 
700 1 |a Arakawa, T  |e verfasserin  |4 aut 
700 1 |a Hara, A  |e verfasserin  |4 aut 
700 1 |a Ohsato, N  |e verfasserin  |4 aut 
700 1 |a Izawa, M  |e verfasserin  |4 aut 
700 1 |a Aizawa, K  |e verfasserin  |4 aut 
700 1 |a Itoh, M  |e verfasserin  |4 aut 
700 1 |a Shibata, K  |e verfasserin  |4 aut 
700 1 |a Shinagawa, A  |e verfasserin  |4 aut 
700 1 |a Kawai, J  |e verfasserin  |4 aut 
700 1 |a Ota, Y  |e verfasserin  |4 aut 
700 1 |a Kikuchi, S  |e verfasserin  |4 aut 
700 1 |a Kishimoto, N  |e verfasserin  |4 aut 
700 1 |a Muramatsu, M  |e verfasserin  |4 aut 
700 1 |a Hayashizaki, Y  |e verfasserin  |4 aut 
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