title: GSM4041604: Pol2 Chia-Pet Control; Mus musculus; ChIA-PET
library_construction_protocol: Whole cell extracts from 500.000 million cells were sonicated. The chromatin extracts containing DNA fragments with an average size of 300-700 bp were immunoprecipitated with anti-RNAPII (Pol2, Clone 8WG16, Ab817, Abcam). Immunoprecipitated DNA was end-repaired, A-tailed, and proximity ligation was performed in the presence of a bridge-linker (5'-ACGCGATATCTTATCTGACT) with T4 DNA ligase. Unligated DNA was digested with exonuclease treatment followed by RNase A, and Proteinase K treatment. DNA was then phenol:chloroform:isoamyl alcohol precipitated and subjected to library preparation. Purified immunoprecipitated DNA was tagmented and amplified with Nextera DNA Flex Library Prep (20018704, Illumina) and Nextera DNA CD Indexes (115055289, Illumina), according to the manufacturer's protocol. Following amplification, the PCR products were purified with AMPure beads (Beckman Coulter) and resuspended in 40 μl Buffer EB (Qiagen). The purified library was size selected (200-700 bp) with a Pippin prep machine. Illumina Hi-Seq 2500 platform at the Genome Tech Core at the Whitehead Institute for Biomedical Research. 100x100 paired-end sequencing.
attributes.tag: GEO Accession
attributes.value: GSM4041604