GSM2837709: U937_rep1_p66Shc; Homo sapiens; OTHER
HUVEC or U937 cells (5*10^6) were cross-linked with formaldehyde and treated with lysis buffer, nuclei were digested with DpnII (NEB) and ligated with T4 DNA ligase (NEB). Protein K was used for de-cross-link, and phenol-chloroform was used for the genomic DNA extraction. Subsequently, the extracted genomic DNA was digested with Csp6I (NEB) in a second round, and the DNA was ligated again with T4 DNA ligase (NEB). Specific primers were designed near the p66Shc promoter. The viewpoint was a 562bp DNA fragment which coordinated at -335bp ~+227bp of p66Shc TSS. Illumina adaptors were included in the primers. PCRs were performed with Expand Long Template PCR System (Roche), PCR products were purified using QIAquick PCR Purification Kit (Qiagen) and the sample concentrations were measured by Qubit dsDNA BR Assay Kit (Thermo Fisher). All samples were sequenced using PE150 program with Illumina Hi-Seq X Ten.
GEO Accession
GSM2837709