title: GSM8438385: male SMG H3K4me1 ChIP; Homo sapiens; ChIP-Seq
library_construction_protocol: Salivary gland samples were dissected and subjected to chemical and enzymatic digestion to generate single cell suspensions which were then cross-linked and processed for downstream applications. For histone ChIP-seq the Chromatin Shearing Optimization Kit-Low SDS (Diagenode) was used with 10 million cells per cell line and 40 cycles sonication in the Bioruptor Plus (Diagenode). After RNase and proteinase K incubation, chromatin shearing was confirmed as a DNA smear between 100-300 bp on a 1.5% agarose gel. Histone ChIP DNA was prepared with the Auto Histone ChIP-seq kit protein G Kit (Diagenode), 2.4 μg H3K27ac, with 3 μg H3K4me1, 2 μg H3K4me3, and 2 μg H3K4me2 of premium polyclonal antibody (Diagenode) per 1 million cells with the SX-8G IP-Star® Compact System (Diagenode). 1% input sample for each cell line was used as a control for background experimental noise. ChIP-sequencing on ilumina HiSeq2500. Libraries were prepared using ThruPLEX DNA-seq kits from Rubicon Genomics. The samples were then 50 bp single-end sequenced on an Illumina HiSeq 2500.