RNA-Seq of human: siControl HAC15 Batch 3
The human adrenocortical carcinoma (HAC15)cells were silenced either using the Silencer Select Negative Control siRNA #2 as control (Control), or Silencer Select Pre-Designed siRNA CYP11B2 (siCYP11B2). HAC15 cells (1000000 cells) were transfected with 0.2 M of siCYP11B2 or Control using the Neon Transfection System. Experiment was performed  3 times independently in which each treatment/ transfection was performed in triplicate.Total DNA-free RNA from cells was isolated using the PureLink RNA Mini Kit. RNA sequencing (RNA-seq) was conducted on representative samples derived from Control and siCYP11B2-transfected cells, obtained from three independent biological replicates. cDNA library constructions were performed and paired-end (PE100) reads were sequenced commercially using the Sequencing Platform DNBseq (Beijing Genomics Institute (BGI, https://www.bgi.com). Clean raw sequences (Phred+33) were obtained through a series of data processing using the SOAPnuke software developed by the sequencing company. Dr. Tom, a web-based solution designed by the BGI for the convenient analysis, visualization and interpretation of the RNA data, was used to identify the differential expressed genes (DEG) and to perform the pathway analysis (https://biosys.bgi.com/#/report/login).    Differential expression analysis was performed by aligning clean reads to the reference genome (GCF_000001405.38_GRCh38.p12) and applying a threshold of |logFC|>1 with an adjusted P-value<0.05 to identify significantly differentially expressed genes (DEGs). Average FPKM for siCYP11B2 is 7.48 and Control is 23.64.