title: GSM1903799: Panct1 WT - Odd probes ChIRP-Seq; Mus musculus; OTHER
library_construction_protocol: Following crosslinking, chromatin was sonicated, incubated with Biotin labelled probes overnight and pulled down using Streptavidin beads. DNA was then extracted from the beads, treated with RNAse A and RNAse H, proteinase K and finally purified. Immunoprecipitated fragments were end-repaired with NEBnext End Repair Module (NEB) and purified by using Agencourt Ampure XP - beads (Beckman Coulter) and A-tailed using the NEBnext dA-Tailing Module according to manufacturers’ instructions. After purification, adaptors were ligated (Adaptor-Oligo 1: 5'ACACTCTTTCCCTACACGACGCTCTTCCGATCT-3', Adaptor-Oligo 2: 5'-P-GATCGGAAGAGCACACGTCTGAACTCCAGTCAC-3') by using 1x NEBnext Quick Ligation Buffer (NEB), 10x excess of DNA Adaptors, 5 µl Quick T4 DNA Ligase (NEB) at 50 µl total volume. Large-scale amplification of library constructs was performed by using the PCR Enrich Adaptor Ligated cDNA Library module (NEB).
attributes.tag: GEO Accession
attributes.value: GSM1903799