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A cell-sorting-based protocol for cell cycle small-scale ChIP sequencing.

STAR protocols | 2022

Classic approaches to characterizing cell cycle leverage chemicals or altered nucleotide pools, which could impact chromatin states at specific phases of the cell cycle. Such approaches could induce metabolic alterations and/or DNA damage, which could reshape protein recruitment and histone modifications. In this protocol, we describe ways to fix and sort cells across the cell cycle based on their DNA content. We further detail immunoprecipitation and library preparation, allowing analysis of the epigenome by chromatin immunoprecipitation sequencing (ChIP-seq) for small numbers of cells. For complete details on the use and execution of this protocol, please refer to Van Rechem et al. (2021).

Pubmed ID: 35310076 RIS Download

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HEK293T (tool)

RRID:CVCL_0063

Cell line HEK293T is a Transformed cell line with a species of origin Homo sapiens (Human)

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Histone H3 (di methyl K36) antibody - ChIP Grade (antibody)

RRID:AB_1280939

This polyclonal targets Histone H3 (di methyl K36) antibody - ChIP Grade

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Histone H3 (di methyl K9) antibody [mAbcam 1220] - ChIP Grade (antibody)

RRID:AB_449854

This monoclonal targets Histone H3 (di methyl K9) antibody [mAbcam 1220] - ChIP Grade

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H3K9me1-human (antibody)

RRID:AB_732929

This polyclonal targets H3K9me1

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H3K36me3-human (antibody)

RRID:AB_306966

This polyclonal targets H3K36me3

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H3K9me3-human,H3K9me3-mouse (antibody)

RRID:AB_306848

This polyclonal targets H3K9me3

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H3K27ac-mouse (antibody)

RRID:AB_2561016

This polyclonal targets H3K27ac

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H3K4me2-human (antibody)

RRID:AB_732924

This monoclonal targets H3K4me2

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