title: Detecting expressed genes using CAGE.
abstract: 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.
authors.affiliation: Division of Genomic Technologies, RIKEN Center for Life Science Technologies, RIKEN Yokohama Institute, 1-7-22 Suehiro-cho, Tsurumi-ku, Yokohama, Kanagawa, 230-0045, Japan.
authors.forename: Mitsuyoshi
authors.initials: M
authors.lastname: Murata
authors.forename: Hiromi
authors.initials: H
authors.lastname: Nishiyori-Sueki
authors.forename: Miki
authors.initials: M
authors.lastname: Kojima-Ishiyama
authors.forename: Piero
authors.initials: P
authors.lastname: Carninci
authors.forename: Yoshihide
authors.initials: Y
authors.lastname: Hayashizaki
authors.forename: Masayoshi
authors.initials: M
authors.lastname: Itoh
mesh_terms: D000469:Alkaline Phosphatase / Q000378:metabolism; D000818:Animals; D001483:Base Sequence; D019921:Biotinylation / Q000379:methods; D018076:DNA, Complementary / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D005090:Exodeoxyribonucleases / Q000378:metabolism; D020869:Gene Expression Profiling / Q000379:methods*; D015723:Gene Library; D059014:High-Throughput Nucleotide Sequencing / Q000379:methods*; D006801:Humans; D051379:Mice; D011401:Promoter Regions, Genetic; D012313:RNA / Q000235:genetics* / Q000378:metabolism; D048348:Reverse Transcription; D012259:Ribonuclease, Pancreatic / Q000378:metabolism; D024363:Transcription Initiation Site
chemical_list: D018076:DNA, Complementary; D012313:RNA; D005090:Exodeoxyribonucleases; C028450:exodeoxyribonuclease I; D012259:Ribonuclease, Pancreatic; D000469:Alkaline Phosphatase