GSM8329652: eCLIP_IgG_rep2; Homo sapiens; OTHER
TOP1-RNA complexes were separated by SDS-PAGE, transferred to a nitrocellulosemembrane, and RNA was extracted from the membrane using proteinase K (NEB). 3' RNA adaptor was ligated onto the RNA with T4 RNA ligase. RNA was reverse transcribed with Superscript IV (Invitrogen), free primer was removed (ExoSap-IT, Affymetrix) and a 3' DNA adaptor was ligated onto the cDNA product with T4 RNA ligase (NEB). Complementary DNA was amplified by PCR using Q5 Master Mix (NEB) and purified before Illumina sequencing. The sequencing was done using Illumina NextSeq 1000 with a read length configuration of 75bp for 40M reads per sample.