GSM8586337: ilmn_Colo320_rep1; Homo sapiens; RNA-Seq
Cells were collected and lysed in TRIzol reagent (ThermoFisher). The lysate was mixed with chloroform and phase-separated in a phaselock gel tube heavy (Quantabio) by centrifugation. The aqueous phase containing RNA was then mixed with one volume of 100% ethanol, and column-purified using Zymo QuickRNA Miniprep kit with in-column DNase treatment per the manufacturer's protocol. RNA integrity (RIN  ≥ 9) was confirmed using the 4200 Tapestation System (Agilent) prior to library construction. Long-read RNA-seq libraries were constructed using the NEBNext Single Cell/Low Input cDNA Synthesis & Amplification Module, PacBio Iso-Seq Express Oligo Kit and SMRTbell express template prep kit 3.0 according to the manufacturer's protocol. Short-read RNA-seq libraries were prepared using the SMARTer Stranded Total RNA-seq kit v3 (TaKaRa Bio) following the manufacturer's instructions.