title: GSM9375662: RNA_WT_aYFP_PolyAMinus_rep3; Mus musculus; RNA-Seq
library_construction_protocol: Total RNA was collected from 500,000 cells lysed in Trizol (Sigma: T9424) and extracted by 1-Bromo-3-Chloropropane. RNA was treated with Turbo Dnase to remove gDNA contamination, and extracted by phenol:chloroform:isoamyl alcohol (25:24:1) For RNA-seq libraries, 500 ng of Turbo Dnase treated total RNA was depleted of rRNA using the NEBNext rRNA Depletion Kit (E6310) following manufacturer instructions. PolyA selection was performed usign the NEBNext Poly(A) mRNA Magnetic Isolation Module (New England Biolabs: E7490) following manufacturer instructions. Stand-specific cDNA as synthesised and each sample uniquely indexed for multiplex Illumina sequencing using the NEBNext Ultra II Directional RNA Library Prep Kit (New England Biolabs: E7760). Both Poly(A)+ and Poly(A)- RNA-seq libraries were sequenced.