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

被引：19

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

来源：

NUCLEIC ACIDS RESEARCH | 2004年 / 32卷 / 01期

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.

引用

页数：6