RNA-seq of Homo sapiens: Antibody-based biotinylated RNA in HeLa cell
After biotinylation and avidin pull-down, the bound protein-RNA complex was eluted and reverse-crosslinked by incubation for 1 h at 55C in TE buffer containing 1% SDS, 0.2 M NaCl and proteinase K. The pulled-down RNAs were isolated by Trizol LS (Invitrogen, 10296028) and ethanol precipitation. cDNA library was prepared by NEBNext Ultra Directional RNA Library Prep Kit for Illumina (New England Biolabs).