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.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Alkaline Phosphatase / metabolism
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Animals
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Base Sequence
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Biotinylation / methods
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DNA, Complementary / genetics*
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DNA, Complementary / isolation & purification
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DNA, Complementary / metabolism
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Exodeoxyribonucleases / metabolism
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Gene Expression Profiling / methods*
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Gene Library
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High-Throughput Nucleotide Sequencing / methods*
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Humans
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Mice
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Promoter Regions, Genetic
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RNA / genetics*
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RNA / metabolism
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Reverse Transcription
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Ribonuclease, Pancreatic / metabolism
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Transcription Initiation Site*
Substances
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DNA, Complementary
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RNA
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Exodeoxyribonucleases
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exodeoxyribonuclease I
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Ribonuclease, Pancreatic
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Alkaline Phosphatase