H3K9me3 Nucleosome Sequencing

Mono nucleosomes were prepared from cultured CD8 T cells by micrococcal nuclease digestion as described (30 (link)). H3K9me3-modified nucleosomes were immunoprecipitated using anti-H3K9me3 (Abcam 8898) conjugated with Protein G magnetic beads (Life Technologies). For genome-wide analysis, purified DNA from precipitated nucleosomes was sequenced with a HiSeq 2500 sequencer (Illumina) with a 50-bp single end read option as described (23 (link)).

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Verbaro D.J., Sakurai N., Kim B., Shinkai Y, & Egawa T. (2018). The histone methyltransferase G9a is required for silencing of helper T lineage-associated genes in proliferating CD8 T cells. Journal of immunology (Baltimore, Md. : 1950), 200(12), 3891-3896.

Publication 2018

anti-H3K9me3 Protein G magnetic beads

Cd8 t cells Digestion Genome Micrococcal nuclease Nucleosomes Protein g

Corresponding Organization : Washington University in St. Louis

Other organizations : RIKEN

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Variable analysis

independent variables

Micrococcal nuclease digestion

dependent variables

H3K9me3-modified nucleosomes
Purified DNA from precipitated nucleosomes

control variables

Cultured CD8 T cells

controls

Positive control: Anti-H3K9me3 (Abcam 8898) conjugated with Protein G magnetic beads (Life Technologies) used for immunoprecipitation
Negative control: Not explicitly mentioned

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