Improved full-length cDNA production based on RNA tagging by T4 DNA ligase

AUTOR(ES)

Clepet, Christian

FONTE

Oxford University Press

RESUMO

Second-strand cDNA priming is a central problem for full-length characterization of transcripts. A new strategy using bacteriophage T4 DNA ligase and partially degenerate adapters is proposed for grafting a sequence tag to the end of polyribonucleotides. Based on this RNA tagging system and previously described protocols, a new method for full-length cDNA production has been implemented. Validation of the method is shown in Arabidopsis thaliana by the construction of a full-length cDNA library and the analysis of 154 clones and by 5′-RACE–PCR run on a documented experimental system.

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