GSM5774545: Cal27-CYTOR-KD-3; Homo sapiens; RNA-Seq
Cells were washed with 1 X PBS for 3 times, and RNA was harvested using Trizol reagent. Illumina TruSeq RNA Sample Prep Kit (Cat#FC-122-1001) was used with 1-3 ug of total RNA for the construction of sequencing libraries. mRNA was captured from approximately 1μg of total RNA using VAHTSTM mRNA Capture Beads. Next, mRNA-seq library was prepared with RNA-seq Library Prep Kit for Illumina  (Geneseed, Guangzhou, China). Briefly, the mRNA was fragmented and then first strand and directional second strand synthesis were performed. Then the A tailing and adapter ligation were performed with the purified cDNA. Finally, the purified, adapter-ligated DNA was amplified. The library quality and concentration was assessed by utilizing a DNA 1000 chip on an Agilent 2100 Bioanalyzer.