GSM7994218: EP000004A01_7_12.711864CF_TC00503572_K04; Homo sapiens; RNA-Seq
Six hours after water extract treatment, HTTr assay plates were removed from the incubator and media in each assay well drained to a residual volume of 10 µL. A 10 µL volume of 2× BioSpyder lysis buffer was dispensed into each assay well. Then, 20 µL volumes of Universal Human Reference RNA (UHRR), Human Brain Reference RNA (HBRR), bulk lysates from MCF7 cells treated with 0.5% DMSO (BL DMSO) or 1 µM trichostatin A (BL TSA), and lysis buffer blans were dispensed into the first column of each assay plate. The plates were sealed with an adhesive aluminum plate cover and incubated at room temperature, protected from light, for 30 min. The plates were then stored at -80°C prior to shipment to BioSpyder, Inc. Samples were analyzed by BioSpyder using a custom-attenuated version of the TempO-Seq human whole transcriptome version 1 (hWTv1) assay, which includes 21,111 probes covering 19,287 genes. In brief, 2 µL of each lysate was hybridized with 2 µL of detector oligos from the hWTv1 assay using the following thermal cycler protocol: 10 min at 70°C, followed by gradual decrease to 45°C over 49 min, terminating with 45°C incubation for 16-24 hours. Excess oligos were then removed via nuclease digestion (90 min at 37°