title: Illumina HiSeq 4000 paired end sequencing; RNA-seq of early- and late-passage human mesenchymal stem cells (5 donors, donor-matched) with and without treatment with 2 hours of 42 degrees C heat shock
design: RNA-seq of early- and late-passage human mesenchymal stem cells (5 donors, donor-matched) with and without treatment with 2 hours of 42 degrees C heat shock
library_construction_protocol: Early and late passage human mesenchymal stem cells were heat shocked (42°C, 2 hour, 5% CO2) and compared with controls kept at 37°C, 5% CO2. Following treatment, cells in culture flasks were washed with PBS, lysed in-flask, and material was collected using a cell scraper. Cells were lysed and RNA was extracted using the RNeasy Mini kit (Qiagen) and an on-column DNA digest performed using the RNase-Free DNase kit (Qiagen). The RNA was eluted in water and the quality and quantity assessed using a NanoDrop ND-2000 spectrometer (Thermo Fisher). TruSeq® Stranded mRNA assay (Illumina, Inc.) according to the manufacturer's protocol.