title: GSM5520136: RNA-seq_N3; Homo sapiens; RNA-Seq
library_construction_protocol: Total RNA product was purified using RNeasy Mini Kit (QIAGEN, Germany) and run-on Agilent Bioanalyzer 4200 to assess RNA integrity. The total RNA‐seq library preparation from 1 μg of total RNA was performed with TruSeq RNA Sample Prep Kit (Illumina, USA) according to the manufacturer's instructions. The ATAC-seq assay was performed using 0.1 million cells isolated from normal or tumor tissues were lysed in 100μL lysis buffer per sample (10 mM Tris-HCl at pH 7.4, 10 mM NaCl, 3 mM MgCl2 and 0.1% IGEPAL CA-630) for 10 min on ice to prepare the nuclei briefly. Immediately After 10 min, nuclei were spun for 5 min at 500 x g to remove the supernatant. Then nuclei were incubated at 37 °C with the tagmentation buffer and Tn5 transposome (Vazyme Biotech, China) for 30 min. After the 30 min, ended buffer was added directly into the reaction to quit the tagmentation. Products which were inputted to PCR were performed to amplify the library for 11 cycles using the following PCR conditions: 72 °C for 3 min; 98 °C for 30 s; and thermocycling at 98 °C for 15 s, 60 °C for 30 s and 72 °C for 3 min; following by 72 °C 5 min. After the PCR reaction, DNA libraries were purified with DNA Clean Beads (Vazyme Biotech, China) to remove invalid primer dimmers and quantitate used Qubit 2.0 Fluorimeter (Invitrogen, USA) to quantitate. All eligible DNA libraries were sequenced on the Illumina HiSeq X Ten with 150 bp paired-end reads.