Detecting Expressed Genes Using CAGE
Detecting Expressed Genes Using CAGE
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DOI：
10.1007/978-1-4939-0805-9_7
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发表时间：
2014-01-01
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通讯作者：
Itoh, Masayoshi
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作者：
Murata, Mitsuyoshi;Nishiyori-Sueki, Hiromi;Itoh, Masayoshi
基因表达帽分析 (CAGE) 提供 RNA 表达的准确高通量测量。通过使用CAGE方法对转录本5'末端进行大规模分析，不仅可以确定转录起始位点，还可以预测启动子区域。在这里，我们提供了用于构建 Illumina 下一代测序仪 (nAnT-iCAGE) 的非扩增非标记 CAGE 库的协议。我们排除了常用的 PCR 扩增和限制性内切酶切割，以消除任何潜在的偏差。结果，我们实现了偏差较小的简单制备过程。
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.