GSM2481420: 16HBE14o- DNase-seq; Homo sapiens; DNase-Hypersensitivity
DNase-seq: genomic DNA was digested with different concentrations of DNaseI (NEB) and purified for library construction (Boyle et al. Cell, 2008). ChIP-seq: Cells were lysed in lysis buffer (5mM PIPES pH 8.0, 85mM KCL, 0.5% NP-40, 1x Protese Inhibitor Cocktail (Roche)). Nuclei were isolated by centrifugation, lysed in RIPA buffer (1x PBS, 1% NP-40, 0.5% sodium deoxycholate, 0.1% SDS, Protease Inhibitor Cocktail), and sonicated. Protein-DNA complexes were isolated with the histone antibodies. DNase-seq: libraries were prepared by Drs A. Safa and G. Crawford. ChIP-seq: DNA ends were blunted using T4 DNA polymerase, Klenow DNA polymerase, and T4 polynucleotide kinase. Next, DNA was incubated with Klenow exo- to add 5’ adenine overhangs. Multiplex adaptors were then ligated to the ends and converted to dsDNA using 5 cycles of PCR. DNA was size-selected to contain fragments between 200 and 300bp and PCR amplified for 5 cycles. DNA was purified with 1.2x AMPsureXP beads. Sequencing was preformed on an Illumina Hi-Seq.
GEO
GSM2481420
GEO Accession
GSM2481420