Illumina HiSeq 2500 sequencing; RNA Sequencing of CDC27 downregulation in SK-N-SH cells
RNA Sequencing of CDC27 downregulation in SK-N-SH cells
Cell lines used in our study were purchased from Procell Life Science & Technology (Wuhan, China), and were cultured in standard medium containing 10% fetal bovine serum. Cell lines authentication were performed using short tandem repeat DNA profiling, and mycoplasma detection was regularly performed using a PCR-based method. All cell lines were cultured in a humidified atmosphere with 5% CO2 at 37C Total RNA was isolated using the Trizol reagent (Invitrogen), and conversion to cDNA was completed using the GoScript Reverse Transcription System (Promega). The relevant mRNA expression of target genes was quantified by Real-time PCR using a SYBR Green PCR Kit (Invitrogen) normalized to GAPDH. Sequenced reads were trimmed for adaptor sequence, and masked for low-complexity or low-quality sequence, then mapped to hg19 whole genome using Tophat2 v2.0.13 with parameters --read_mismatches 2 --read_gap_length 2 --read_edit_dist 2