Cap analysis of gene expression (CAGE) provides accurate high-throughput measurement of RNA expression. By the large-scale analysis of 5′ end of transcripts using CAGE method, it enables not only determination of the transcription start site but also prediction of promoter region. Here we provide a protocol for the construction of no-amplification non-tagging CAGE libraries for Illumina next-generation sequencers (nAnT-iCAGE). We have excluded the commonly used PCR amplification and cleavage of restriction enzyme to eliminate any potential biases. As a result, we achieved less biased simple preparation process.
Cap analysis of gene expression (CAGE) RNA expression Transcription start site (TSS) Promoter Next-generation sequencing (NGS)
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This work was funded by a Research Grant from the Japanese Ministry of Education, Culture, Sports, Science and Technology through the Cell Innovation Project and for the RIKEN Omics Science Center to Y.H.
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