GSM7329979: H1_D2_SS2_ATCACG; Homo sapiens; RNA-Seq
Total RNA was harvested from TeloHAEC (parental or CRISPRi lines) by Qiagen RNeasy kit, washed with 350 ul of buffer RW1 and treated for 20 mins at room temperature with 10 ul Purelink DNAse (InVitrogen 12185010)  in 80 ul of 1x buffer, with subsequent steps according to the supplier's method. For shear stress (flow) versus static cells, RNA was instead harvested with 300 ul of Trizol and extracted with 60ul of chloroform. After addition of 1 volume 70% ethanol, RNA was loaded onto a Qiagen RNeasy spin column, with subsequent steps as per the Qiagen RNeasy protocol, above. mRNA was purified from 400ng to 1 ug of total RNA using the NEBNext Poly(A) mRNA Magnetic Isolation module (NEB). NEBNext Ultra II RNA Library Kit for Illumina (NEB), according to manufacturer's protocol, and index primers specific for each library