title: RNA seq of placental organoids generated from CT29 cell lines treated with PRKCZ-targetting siRNA versus control
design: RNA seq of placental organoids generated from CT29 cell lines treated with PRKCZ-targetting siRNA versus control
library_construction_protocol: CT29 cell lines were grown and treated with either PRKCZ-targetting siRNA or a scrambled control. The cell lines were used to generate placental organoids. CT 29 lines were maintained on 5 Âµg/mL collagen IV coated plates and using human TSC culture medium in a humidified incubator at 37 degrees and 5% CO2. Organoids were generated using a rotatory cell culture system. CT29 knockdowns were achieved utilizing PRKCZ-targetting siRNA that was transfected into the cell line using Opti-MEM I Reduced Serum Medium and Lipofectamine LTX Reagent with PLUS reagent.  Placental organoids were pelleted and resuspended in trizol before being mechanically dissociated utilizing a tissue lyser. RNA was extracted by utilizing Invitrogen PureLink RNA Mini Kit. Bulk seq libraries were constructed using the Illumina Stranded mRNA Prep Kit.