Improved full-length cDNA production based on RNA tagging by T4 DNA ligase
Nucleic Acids ResearchPublished 2 January 2004Open access
Christian Clépet
Citations42
SJR quartileQ1
SJR score7.78
SNIP4.71
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TL;DR
A new method for full-length cDNA production has been implemented using bacteriophage T4 DNA ligase and partially degenerate adapters for grafting a sequence tag to the end of polyribonucleotides.
Abstract
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.
Keywords
Biochemistry, Genetics and Molecular BiologyEnvironmental Science
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