GSM1643925: 87_4; Mus musculus; ChIP-Seq
Chromatin was sheared to 200-400bp, incubated with H3K4me1/2 antibody (abcam 32356) overnight at 4C on a rocker, antibody chromatin complex was captured by ProteinA agarose beads, washed with low salt, high salt, LiCl, and TE buffer, eluted and then de-crosslinked at 65C with 0.3M NaCl final for at least 6 hr.  Final chormatin was purified using Qiagen PCR purification columns.  RNA was purified using the RNAeasy Plus Kit (QIAGEN) that included a genomic DNA elimination step. DNA was purified using PCR purification columns (QIAGEN). RNA-seq libraries were generated using Illumina’s TruSeq RNA sample Prep Kit v2. ChIP-seq libraries were prepared using 10ng of DNA and Illumina’s TruSeq ChIP sample prep. Libraries were validated using the Agilent Technologies 2100 Bioanalyzer and Quant-iT dsDNA HS Assay (Life Technologies).