GSM8677951: KG1-shUCP2#3; Homo sapiens; RNA-Seq
KG1 cell suspension were washed twice with ice cold PBS and DNA was extracted using DNeasy Blood & Tissue Kit (#69504, Qiagen) per instruction manual DNA samples were end-repaired, A tailed, and adaptor ligated using TruSeq Nano DNA Sample Prep Kit (#FC-121-4002, Illumina) Sequencing library was prepared by converting 10 ng DNA of each sample to phosphorylated blunt-end fragment. An 'A' base was added to the 3' end of the blunt phosphorylated DNA fragments, and Illumina's genomic adapters were ligated to the A tailed DNA fragments prior to subject to PCR amplification to enrich ligated fragments. The enriched product of ~200-1500 bp was size selected using AMPure XP beads. The library was denatured with 0.1M NaOH to generate single-stranded DNA molecules, and loaded onto channels of the flow cell at 8pM concentration, amplified in situ using Cluster Kit.