Transcriptome Analysis of the Anti-Proliferative Effects of Ginsenoside Rh3 on Colorectal Cancer HCT116 Cells

Ginsenoside Rh3 is an important rare protopanaxadiol with a variety of pharmacological and physiological activities including anticancer, however, the underlying mechanism still remains unclear. In this study, transcriptome of untreated and Rh3 treated Colorectal Cancer HCT116 Cells was analyzed by RNA-seq (1) RNA quantification and qualification: Total RNA was extracted using the Trizol method, RNA degradation and contamination was monitored on 1% agarose gels, RNA purity was checked using the NanoPhotometer spectrophotometer (IMPLEN, CA, USA). RNA integrity was assessed using the RNA Nano 6000 Assay Kit of the Bioanalyzer 2100 system (Agilent Technologies, CA, USA). (2) Library preparation for Transcriptome sequencing: A total amount of 1 microgram RNA per sample was used as input material for the RNA sample preparations. Sequencing libraries were generated using NEBNext UltraTM. RNA Library Prep Kit for Illumina (NEB, USA) following manufacturer recommendations and index codes were added to attribute sequences to each sample. (3) Clustering and sequencing: The clustering of the index-coded samples was performed on a cBot Cluster Generation System using TruSeq PE Cluster Kit v3-cBot-HS (Illumia) according to the manufacturer's instructions. After cluster generation, the library preparations were sequenced on an Illumina Novaseq platform and 150 bp paired-end reads were generated.