GSM5491991: PRO-seq in Human K562 cells, rep2; Homo sapiens; OTHER
The cells were harvested by centrifugation and washed twice with PBS. Nuclei were isolated and run-on reactions were performed with biotinylated nucleotides as described by Kwak H, Fuda NJ, Core LJ, Lis JT. Precise maps of RNA polymerase reveal how promoters direct initiation and pausing. Science 2013 Feb 22;339(6122):950-3. The run-on reaction was conducted in the presence of nuclei from S2 cells, which provides an invariant whole-genome spike-in control between samples. The libraries were generated using TruSeq small-RNA adaptors, amplified 11 cycles, gel-purified and size-selected to 125-400 bp fragments. The libraries were sequenced using NextSeq 500 (Illumina).