Abstract:
:Cap-analysis gene expression (CAGE) provides accurate high-throughput measurement of RNA expression. CAGE allows mapping of all the initiation sites of both capped coding and noncoding RNAs. In addition, transcriptional start sites within promoters are characterized at single-nucleotide resolution. The latter allows the regulatory inputs driving gene expression to be studied, which in turn enables the construction of transcriptional networks. Here we provide an optimized protocol for the construction of CAGE libraries on the basis of the preparation of 27-nt-long tags corresponding to initial bases at the 5' ends of capped RNAs. We have optimized the methods using simple steps based on filtration, which altogether takes 4 d to complete. The CAGE tags can be readily sequenced with Illumina sequencers, and upon modification they are also amenable to sequencing using other platforms.
journal_name
Nat Protocjournal_title
Nature protocolsauthors
Takahashi H,Lassmann T,Murata M,Carninci Pdoi
10.1038/nprot.2012.005subject
Has Abstractpub_date
2012-02-23 00:00:00pages
542-61issue
3eissn
1754-2189issn
1750-2799pii
nprot.2012.005journal_volume
7pub_type
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