Random Fragment pulldown RNA from HEK293T cell lysate
HEK293T cell lysates were incubated with biotin-tagged RNA oligonucleotides (ES30L mimic and random fragment; n=2) and pulled down using Dynabeads M-280 Streptavidin beads. RNA was eluted from the based using TRIzol based extraction. RNA from whole cell lysates were also extracted by acetone using TRIzol (referred to as input in the samples; n=2).Sequencing libraries were prepared using NEBNext Ultra II Directional RNA Library Prep after poly(A) selection from the isolated RNA and sequenced on an Illumina Hiseq 2500 machine.