title: Illumina HiSeq 4000 sequencing; Axon Guidance Receptor ROBO3 Modulates Subtype Identity and Prognosis via AXL-associated Inflammatory Network in Pancreatic Cancer
design: Axon Guidance Receptor ROBO3 Modulates Subtype Identity and Prognosis via AXL-associated Inflammatory Network in Pancreatic Cancer
library_construction_protocol: PANC1 cells were seeded in 6-well plates and transfected with 20 nM siRNA (siControl or siROBO3) using Lipofectamine 2000. RNA was extracted 72 hours after transfdection. Total RNA was extracted from PANC1 cells with TRIzol (Invitrogen) following the manufacturer's recommendations.  mRNA sequencing libraries were generated with the TruSeq RNA Library Prep kits (Illumina) according to the manufacturerâ€™s instructions. The size of the generated libraries was estimated on High Sensitivity DNA chip (Agilent) using a Bioanalyser 2100 (Agilent).