title: GSM4333259: K562D; Homo sapiens; OTHER
library_construction_protocol: All cells were incubated with NAI-N3 at 37°C for 5min. 100 µg of RNA from each samples was depleted of ribosomal RNA (rRNA) using DynabeadsTM mRNA DIRECTTM kit (invitrogen). To sequence the libraries on the Hiseq X system, we designed new linker, reverse transcription (RT) primer and P5, P7 amplification PCR primer. The experimental barcodes for different libraries located on PCR primers rather than on RT primers. The linker of NAI-N3 modification samples: /5rApp/AGATCGGAAGAGCACACGTCT/3ddC/; the linker of DMSO control samples: /5rApp/AGATCGGAAGAGCACACGTCT/3Biotin/; RT primer: /5phos/DDDNNNNNNNNNNAGATCGGAAGAGCGTCGTGGA/iSp18/GGATCC/iSp18/CAGACGTGTGC, D = A/G/T and N = A/T/G/C are used to discriminate PCR duplicates; P5-Solexa PCR primer: 5'-AATGATACGGCGACCACCGAGATCTACACTCTTTCCCTACACGACGCTCTTCC-3'; P7-Solexa PCR primer: 5'-CAAGCAGAAGACGGCATACGAGAT CGAGGCTGGTGACTGGAGTTCAGACGTGTGCTCTTCCGATCT-3'. The underline “CGAGGCTG” is the specific experimental barcode to distinguish different libraries. Libraries of all cell lines were sequenced on Hiseq X system to approximately 200 million reads per replicate.
identifiers.namespace: GEO
identifiers.id: GSM4333259