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ChIP-seq and RNA-seq methods to study circadian control of transcription in mammals.

Methods in enzymology (2015-02-11)
Joseph S Takahashi, Vivek Kumar, Prachi Nakashe, Nobuya Koike, Hung-Chung Huang, Carla B Green, Tae-Kyung Kim
RESUMEN

Genome-wide analyses have revolutionized our ability to study the transcriptional regulation of circadian rhythms. The advent of next-generation sequencing methods has facilitated the use of two such technologies, ChIP-seq and RNA-seq. In this chapter, we describe detailed methods and protocols for these two techniques, with emphasis on their usage in circadian rhythm experiments in the mouse liver, a major target organ of the circadian clock system. Critical factors for these methods are highlighted and issues arising with time series samples for ChIP-seq and RNA-seq are discussed. Finally, detailed protocols for library preparation suitable for Illumina sequencing platforms are presented.

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Sigma-Aldrich
Actinomycin D, from Streptomyces sp., ~98% (HPLC)
Sigma-Aldrich
Anticuerpo anti-ARN polimerasa II subunidad B1 (fosfo-CTD Ser-5), clon 3E8, clone 3E8, from rat