title: Illumina HiSeq 1500 sequencing; PPARβ/δ ligand response in monocyte derived macrophages
design: PPARβ/δ ligand response in monocyte derived macrophages
library_construction_protocol: Cells were isolated on June 06th 2013 from healthy human donor blood of unknown gender via LSM1077 gradient centrifugation. Adherent cells were positivly selected on primaria 6 well dishes (3x10^6 monocytes per well in 2 ml PBS (PAA)) followed by 6 washing steps with PBS. Finally the monocytes were cultured for 6d in humidified incubator at 37 °C, 5 % CO2 in 2ml RPMI medium supplemented with 10 % FCS (both PAA). Cells were treated with control (DMSO), L165 1 µM final concentration or PTS-264 1 µM final concentration for 1d. Cells were lysed in Peqgold (TRIzol-like solution based on phenol and chloroform). RNA was digested with DNase (MN) and purified on a Qiagen Minelute column. Illumina-compatible libraries were made using the Epicentre Scriptseq human/mouse/rat low input Gold kit.
attributes.tag: Experimental Factor: compound
attributes.value: PTS264
attributes.tag: Experimental Factor: dose
attributes.value: 1 micromolar
attributes.tag: LIBRARY_STRAND
attributes.value: first strand