GSM8193340: B16F10 cells, no treatment, rep2; Mus musculus; RNA-Seq
Total RNA was isolated and purified from B16F10 cells using TRIzol (Invitrogen) according to the manufacture. The concentration of RNA was checked using Qubit (Invitrogen), and the integrity was assessed using Bioanalyzer 2100 (Agilent). 1 μg total RNA was used for poly (A) RNA purification using Dynabeads Oligo (dT) 25-61005 (Thermo Fisher) with two rounds of purification. the sequencing library was constructed through PCR, with the program set to 3 stages as following: denaturation at 95 °C for 3 min (first stage); denaturation at 98 °C for 15 secs, annealing at 60 °C for 15 secs, and extension at 72 °C for 30 secs (second stage, run 8 cycles); and final extension at 72 °C for 5 min. At last, the sequencing library was sequenced using illumine Novaseq™ 6000 (LC-Bio Technology CO., Ltd., Hangzhou, China) with the model of 2 × 150 bp paired-end sequencing (PE150).