title: NextSeq 500 paired end sequencing; RNAseq of 6 genome edited human RPE1 cell lines (compared to the parental control) disrupting the MIA3 gene.
design: RNAseq of 6 genome edited human RPE1 cell lines (compared to the parental control) disrupting the MIA3 gene.
library_construction_protocol: Samples were prepared in triplicate (3 x 10cm dishes per sample) until confluent. Cells were rinsed with PBS, trypsinized and washed in 5ml PBS before resuspending in 600µl RLT buffer containing 2M DTT.  RNA was extracted using an RNeasy® Mini Kit (Therm FIsher). RNA concentration was verified using a nanodrop, diluted samples in RNAse-free water to concentrations below 250ng per µl  and snap froze them in liquid nitrogen and stored them at -80C until library preparation. 100ng of total RNA was taken into the Illumina TruSeq Stranded Total RNA with Illumina Ribo-Zero Plus rRNA Depletion kits. All instructions in manufacturer's protocols were followed for the library preparation and sequencing steps.