GSM6645038: anti MacroH2A1.1 Chromatin Immunoprecipitation, MEFs; Mus musculus; ChIP-Seq
Nuclei extraction protocol as described in Ford et at. 2014 (Ford E, Nikopoulou C, Kokkalis A, Thanos D. A method for generating highly multiplexed ChIP-seq libraries. BMC Res Notes. 2014 May 22;7:312. doi: 10.1186/1756-0500-7-312. PMID: 24885602; PMCID: PMC4048252) 1-15 ng of “ChIPped” DNA was blunt-ended by End Repair Enzyme Mix (T4 DNA Polymerase, Klenow Fragment, T4 DNA Polynucleotide kinase), followed by “A”-tailing of 3΄-ends and ligation with the annealed TruSeq Adapters (Illumina). Conversion of the Y-shaped adapters to dsDNA occurred prior to the library size selection through 2.5% Metaphor/SeaKem LE (3:1 ratio) agarose gel electrophoresis. Each library was purified using miniElute columns (Qiagen) and then subjected to pre-amplification. The final quantification of DNA libraries was carried out according to the Quantification Standards of Illumina on an Agilent Technologies 2100 Bioanalyzer (Agilent Technologies).