Dataset features

Specifications


Application: ChIP-on-chip analysis
Number of samples: 8
Release date: Sep 26 2014
Last update date: Jan 2 2015
Access: Public
Dataset link Initiation of DNA replication from non-canonical sites on an origin-depleted chromosome

Experimental Protocol


In this study, we aimed to establish an independent system to investigate how an origin-deficient chromosome is replicated. To this end, we systematically deleted seven well-characterized origins on the left arm of S. cerevisiae chromosome VI and analyzed (1) Orc2 localization during G2/M arrest and (2) BrdU incorporation during synchronous release from G1 arrest into S-phase, and compared the results to wild-type cell signals. For Orc2 ChIP-Chip experiments, Orc-bound DNA was isolated from Orc2-2Xlinker-3XFlag epitope-tagged cells arrested in G2/M using antibodies against Flag. For BrdU ChIP-Chip experiments, actively replicating DNA was isolated from cells harboring a single integrated BrdU incorporation vector released synchronously into 200mM HU using antibodies against BrdU. Immunoprecipitated and input (Orc2) or G1 (BrdU) DNA was then amplified and competitively hybridized to high-resolution strand-specific microarrays covering chromosomes III, VI, and XII.

Repositories


GEO

GSE61743

ArrayExpress

E-GEOD-61743

BioProject

PRJNA262013

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Contact


Toshio Tsukiyama

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