GSM2845123: X63/0_cell_non-treated; Mus musculus; RNA-Seq
The cells were harvested by centrifugation, and the total RNA was isolated using a RNeasy Plus Mini kit (QIAGEN) following the instruction manual. Five microgram of total RNA was treated using a GeneRead rRNA Depletion kit (Qiagen) for remove Ribosomal RNA (rRNA) and an RNeasy MiniElute kit (Qiagen) for cleanup. One hundred ng of rRNA-depleted RNA was incubated at 95 ºC for 10 min for fragmentation and was purified by Magnetic Beads Cleanup Module (Life Technologies). The library was constructed with an RNA-seq library kit ver. 2 (Life Technologies) on ABI library builder (Life Technologies) and were barcoded with Ion Xpress RNA-seq BC primer (Life Technologies, 16 PCR cycles). Barcoded library fragment with the size range of 100-200 bp was selected with Agencourt AMPure XP beads (Beckman Coulter, Brea, CA). The emulsion PCR was performed on the OneTouch system using an Ion P1 OT2 200 kit (Life Technologies) to amplify single library fragments onto microbeads and sequencing was performed on an Ion Proton system using the P1 v3 chip (Life Technologies).