GSM5224185: HCC3_MMS_C3_023; Homo sapiens; RNA-Seq
RNA was not extracted from control and treated HepaRG™ cells, as TempO-Seq libraries were constructed directly from cell lysates according tp the TempO-Seq® Assay User Guide (Version 2.1, September 6, 2017). Four hours following the last treatment, the media was aspirated from control (C0) and treated HepaRG™ cells (C1-C5). The cells were then washed with sterile PBS. Following aspiration of the PBS wash, 1X TempO-Seq lysis buffer was added to lyse the cells for 10 minutes at room temperature on a rocking plate. Lysed cells were then frozen at -80 degrees Celsius until library construction. The TempO-Seq® Assay User Guide (Version 2.1, September 6, 2017) was used to build the TempO-Seq libraries for panels with less than 5,000 probes using the TempO-Seq Reagent Kit with HumanTox+Surrogate Panel DO Kit in a 96-well format). Index Set A was used for plate 1, Index Set B for plate 2 and Index Set C for plate 3 (288 samples in total). Following amplification, libraries were pooled and purified using the Macherey-Nagel NucleoSpin Gel and PCR Cleanup Kit with modifications outlined in the TempO-Seq® User Guide. Pooled, purified libraires were quantified using the KAPA Biosystems qPCR library quantification kit and diluted to 4 nM for denaturation and dilution for sequencing. Human TempO-Seq® Tox+ Surrogate (S1500+) kits - Index Sets A (plate 1), B (plate 2) & C (plate 3); sequenced using an Illumina NextSeq500.
GEO Accession
GSM5224185