H3K4me3 and H3K27me3 ChIP-seq

ChIP-seq was performed as described previously^{19 (link),46 (link)}. Briefly, crosslinked chromatin was fragmented by Micrococcal nuclease digestion and immunoprecipitated with anti-H3K4me3 or anti-H3K27me3 antibodies (Abcam, Cambridge, MA). After ChIP, the crosslinking was reversed and DNA fragments were purified. The sequencing library was constructed using Illumina’s TruSeq sample preparation kit according to the provided instruction. The samples before immunoprecipitation served as input controls. Sequencing was performed using Illumina Nextseq 500 sequencer.

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Yang X., Bam M., Nagarkatti P.S, & Nagarkatti M. (2019). Cannabidiol Regulates Gene Expression in Encephalitogenic T cells Using Histone Methylation and noncoding RNA during Experimental Autoimmune Encephalomyelitis. Scientific Reports, 9, 15780.

Publication 2019

anti-H3K4me3 TruSeq sample preparation kit Nextseq 500 sequencer

Anti antibodies Chip Chip seq Chromatin Digestion H3k4me3 Immunoprecipitation Library Micrococcal nuclease

Corresponding Organization :

Other organizations : University of South Carolina

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Variable analysis

independent variables

Crosslinked chromatin fragmentation by Micrococcal nuclease digestion
Immunoprecipitation with anti-H3K4me3 or anti-H3K27me3 antibodies

dependent variables

DNA fragments obtained after ChIP

control variables

Samples before immunoprecipitation (input controls)

controls

Positive control: Anti-H3K4me3 antibody
Positive control: Anti-H3K27me3 antibody
Negative control: Samples before immunoprecipitation (input controls)

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