alias: E-MTAB-7176:Flow-2_s
title: NextSeq 500 sequencing; MDA-MB-231/1833 bone-metastatic breast cancer cell gene expression after conditioning in conditioned medium from HUVECs conditioned in conditioned medium from static and flow-stimulated MLO-Y4 osteocytes
design: MDA-MB-231/1833 bone-metastatic breast cancer cell gene expression after conditioning in conditioned medium from HUVECs conditioned in conditioned medium from static and flow-stimulated MLO-Y4 osteocytes
library_construction_protocol: HUVECs grown to 100% confluency in 6-well plates were conditioned in 50% MLO-Y4 CM and 50% fresh HUVEC media for 24 hours. The medium was then replaced with 100% fresh HUVEC medium to ensure that there is no leftover osteocyte-secreted factor, and medium was collected from the conditioned HUVECs (HUVEC CM) after 12 hours. MDA-MB-231/1833 bone-metastatic breast cancer cells grown to confluency in 6-well plates were subsequently conditioned in 50% HUVEC CM and 50% fresh MDA-MB-231/1833 media for 24 hours. RNA content was isolated from the conditioned MDA-MB-231/1833 cells.  RNA content was isolated using the RNeasy Mini Kit (74104; Qiagen, Venlo, Limburg, Netherlands) and treated with the DNA-free™ Kit (AM1906; Life Technologies). Library construction was done at The Donnelly Sequencing Centre (University of Toronto). The library was prepared with the TruSeq Stranded mRNA Library Prep Kit (Illumina®).
identifiers.namespace: UNIVERSITY OF TORONTO
identifiers.id: E-MTAB-7176:Flow-2_s