GSM8354691: HBEC-Vector Twist1F187G-Vector HRasG12V-EGFP #2; Homo sapiens; RNA-Seq
Total RNA was isolated using TRIzol™ RNA Isolation Reagent (#15596018, ThermoFisher Sci.) protocol and cleaned with RNeasy mini kit (#74104, Qiagen) according to the manufacturer's protocol. RNA sequencing was performed by Genewiz-Azenta (https://www.genewiz.com/, (RRID: SCR_003177)) from total RNA isolated from HBEC RNA sequencing was performed by Genewiz-Azenta (https://www.genewiz.com/): Library preparation, Illumina, RNA with PolyA selection with ERCC spike-in. Library preparation: NEBNext Ultra II RNA Library Prep kit. Briefly, RNA sequencing libraries were prepared using the NEBNext Ultra RNA Library Prep Kit for Illumina using manufacturer's instructions (NEB, Ipswich, MA, USA). mRNAs were initially enriched with Oligod(T) beads. Enriched mRNAs were fragmented for 15 minutes at 94 °C. First strand and second strand cDNA were subsequently synthesized. cDNA fragments were end repaired and adenylated at 3'ends, and universal adapters were ligated to cDNA fragments, followed by index addition and library enrichment by PCR with limited cycles.