Chromatin immunoprecipitation was performed as previously described (Beagan et al. 2016 (link)). Sequencing libraries were prepared with the NEBNext Ultra Library Prep Kit following the manufacturer's protocol for ChIP-seq library preparation, with amplification over 18 PCR cycles using NEBNext Multiplex Oligos for Illumina. The library was confirmed to contain DNA fragments ranging roughly from 250 to 1200 bp by running a High-Sensitivity DNA assay on an Agilent Bioanalyzer and sequenced with 75 cycles per paired-end on the Illumina NextSeq500.
Protocol detail
ChIP-seq Library Preparation Protocol
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A dna
Assay
Chip seq
Chromatin immunoprecipitation
Library
Oligos
Sensitivity
Corresponding Organization :
Other organizations : University of Pennsylvania
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Variable analysis
independent variables
- Chromatin immunoprecipitation
- DNA fragments ranging roughly from 250 to 1200 bp
- Chromatin immunoprecipitation procedure as previously described (Beagan et al. 2016)
- Library preparation with NEBNext Ultra Library Prep Kit
- PCR amplification over 18 cycles using NEBNext Multiplex Oligos for Illumina
- Sequencing with 75 cycles per paired-end on the Illumina NextSeq500
- Positive control: Not specified
- Negative control: Not specified
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