NextSeq 550 sequencing
DiCLIP CATL library from formaldehyde-fixed HUVECs immunoprecipitated with anti-HuR.
DiCLIP libraries were generated from formaldehyde-fixed HUVEC lysates after endogenous RNase trimming, immunoprecipitation with anti-HuR or control IgG, papain-mediated elution of HuR-RNA complexes, de-crosslinking, and RNA purification with the BioLiqX cf-RNA Isolation Kit (HB205A). Libraries were prepared using the BioLiqX Small RNA-Seq Kit (HB202A) with the CATL semi-ligation workflow from sub-nanogram RNA input and sequenced as single-end 75 nt reads on an Illumina NextSeq 550.