title: GSM9232892: RPB1degron_167nM_dTAGv1_1; Homo sapiens; RNA-Seq
library_construction_protocol: Sixteen hours after treatment with dTAGv-1, media was aspirated and cells were harvested by the addition of 1mL Trizol to the plate. To isolate total RNA, one-fifth of chloroform was added to lysates and shaken well, followed by centrifugation (12000g, 5 minutes at 4C). The upper aqueous phase was transferred to MaXtract high-density phase-lock tubes, that were previously prepared according to manufacturer's instructions, and an equal volume of chloroform/isoamyl alcohol (24:1) was added. Lysates were then centrifuged again (12000g, 5 minutes at 4C) and the upper aqueous phase was transferred to a new tube. The RNA was then precipitated through the addition of 1.1X volumes of 100% isopropanol, incubation at room temperature for 20 minutes and centrifugation (12000g, 20 minutes at 4C). RNA pellets were washed once with 85% (v/v) ethanol, and air dried at room temperature before resuspension in Ultrapure DNase/RNase-free distilled water. 2ug total RNA was used for oligo(dT) purification. Sequencing libraries were then constructed using a KAPA mRNA HyperPrep Kit