GSM5453667: cells B total chimeric eCLIP size-matched input, rep 1; Homo sapiens; RIP-Seq
CB replicates of (~5M million cells) and Kasumi-1 cells (~2.5 million cells) were treated with 400mJ of UV using the Hoefer UVC500 UV Crosslinker, harvested in ice cold PBS and pelleted by centrifugation. Cell pellets were flash frozen in liquid nitrogen and stored in -80 degrees until ready to IP with AGO2 bound Dynabeads. Subsequent protocol was followed exactly as detailed in Van Nostrand et al 2016. Sequencing was performed on Illumina Novaseq 6000 paired end reads and data processed through Dr. Yeo's eCLIP pipeline. Libraries were constructed as per the eCLIP protocol using Illumina 50x forward and 70x reverse primers. eCLIP