ChIP-Atlas: SRX1601697

Antigen

library_strategy: ChIP-Seq
library_source: GENOMIC
library_selection: ChIP
library_construction_protocol: The mini-ChIP assays were performed (as described in Adli and Bernstein, 2011) on human neuronal cells that had been cultured for around two weeks. Cells were cross-linked, lysed, sonicated for 3.5 minutes using a Branson 250, and the fragmented chromatin was immunoprecipated overnight with 1ug of H3K27Ac (abcam Cat# ab4728) and H3K27me3 (Millipore Cat# 074490) antibodies. After samples were incubated with Protein A-Sepharose beads for 2 hours they were collected and washed; the DNA was eluted from the beads twice at 65°C for 10 minutes; the eluted chromatin and the "input" samples were reverse cross-linked at 65°C for 5 hours, then Proteinase K digested at 37°C for 2 hours. The ChIP DNA was recovered by standard phenol-chloroform-isoamyl alcohol extraction, then precipitated overnight with ethanol. The ChIP DNA libraries were constructed using ChIP-Seq DNA Sample Prep Kit (Illumina) and sequenced using HiSeq 2000 (Illumina) in the Biopolymers facility at Harvard Medical School. The libraries for RNA-Seq were constructed using the PE RNAseq library kit (Illumina). Strand-specific and paired-end for RNA-Seq, non-strand-specific and single-end for ChIP-Seq.