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

被引:18
作者
Clepet, C [1 ]
Le Clainche, I [1 ]
Caboche, M [1 ]
机构
[1] INRA, CNRS, Unite Rech Genom Vegetale, F-91057 Evry, France
关键词
D O I
10.1093/nar/gng158
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
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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页数:6
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