GSM5195736: TBK1wt_IKBa_shMYC_Dox_rep1; Mus musculus; RNA-Seq
For RNA-seq in vitro experiments, total RNA was isolated by using the RNeasy Mini Kit (Qiagen) including on-column DNaseI digestion. mRNA was isolated by using NEBNext Poly(A) mRNA Magnetic Isolation Module (New England Biolabs). For RNA-seq in vivo experiments, tissue samples were extracted and frozen on dry ice. Total RNA was isolated by using the RNeasy Mini Kit (Qiagen) including on-column DNaseI digestion. mRNA was isolated by using NEBNext Poly(A) mRNA Magnetic Isolation Module (New England Biolabs). cDNA libraries were generated with 1 µg of purified mRNA by using NEBNext Ultra RNA Library Prep Kit for Illumina (New England Biolabs). Size-selection of libraries was performed using Agencourt AMPure XP Beads (Beckman Coulter). The quality, quantity and the size of the cDNA libraries were determined with the Fragment Analyzer (Thermo Fisher Scientific, previously Advanced Analytical Technologies). Libraries were sequenced on a NextSeq500 System (Illumina).