Illumina NovaSeq 6000 paired end sequencing; RNA-seq of HCT116 cells upon downregulation of PINK1
RNA-seq of HCT116 cells upon downregulation of PINK1
Cells were trypsinized, harvested and washed with PBS 1%. Then, the pellet was immediately used for RNA extraction or frozen and stocked at -80Â°C until use. HCT116 cells were grown in DMEM supplemented with 10% Fetal Bovine Serum. About 1.2e6 HCT116 cells were plated on 6 cm plates. The day after cells were overnight infected with SCR or shPINK1 constructs. After 24h, Puromycin was added at 2ug/ml concentration for each plate. After three days of selection, cells were harvested for RNA extraction and sequencing.  RNA extraction was performed using the RNeasy kit (74106) from Qiagen. Ribo minus total RNA stranded kit was used.