title: GSM6365686: SAMPLE 32_MCF7_3D_R2; Homo sapiens; RNA-Seq
library_construction_protocol: Total RNA was extracted from MCF7 and MCF10A cells in both 2D and 3D cultures using RNeasy plus kits (Qiagen, Germany) according to manufacturer's instructions. Briefly, cells grown as monolayer were trypsinized and washed with PBS twice followed by RNA extraction using RNeasy Plus Kit. To collect spheroids, medium was removed from 3D cell culture followed by washing with PBS twice gently. Pre-chilled cell recovery solution (Corning, USA) at the volume of 200 μl were added to 8-well chamber slide (Ibidi, Germany). Matrigel matrix was fully depolymerized, and spheroids were released after 30 minutes incubation at 4°C. Suspended spheroids were collected and washed with PBS twice followed by RNA extraction using RNeasy Plus Kit (Qiagen). The integrity and quantity of RNA were determined with NanoDrop 2000 spectrophotometer (Thermo Fisher Scientific, USA) before sending out for RNA sequencing. Library construction was performed using NEBNext Ultra II RNA Library Prep Kit (Illumina, USA) according to manufacturer's protocols