title: Illumina HiSeq 4000 paired end sequencing; RNASeq of early and late passage (non-senescent and senescent) human mesenchymal stem cells cultured on soft (2 kPa) and stiff (25 kPa) collagen-I coated polyacrylamide gels.
design: RNASeq of early and late passage (non-senescent and senescent) human mesenchymal stem cells cultured on soft (2 kPa) and stiff (25 kPa) collagen-I coated polyacrylamide gels.
library_name: d210_p3_25kPA_p
library_construction_protocol: Early (non-senescent cells) and late (replicative senescent cells) passage human mesenchymal stem cells were cultured on soft (2 kPa) and stiff (25 kPa) PA gels for four days. Following culture, cells were washed with PBS, dissociated using trypsin EDTA, pelleted and frozen at -80Â°C for future use. RNA was extracted from cell pellets 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-1000 spectrometer (Thermo Fisher). Truseq stranded mRNA Assay. Paired end.