GSM3684376: Dolastatin 15; 100 nM treatment at 16H sample 2; Homo sapiens; RNA-Seq
16 hours and 24 hours post drug treatment RNA was extracted using a qiagen kit following the manufacturers protocol for cells. RNA concentration was determined on Qubit® 2.0 Fluorometer (ThermoFisher/Invitrogen, Grand Island, NY), RNA quality was assessed using the Agilent 2100 Bioanalyzer (Agilent Technologies, Inc.). Total RNA with 28S/18S > 1 and RNA integrity number (RIN) ≥ 7 are used for RNAseq library construction.   RNAseq library were constructed using NEBNext® Ultra™ Directional RNA Library Prep Kit for Illumina (NEB, USA) following manufacturer's recommendations. Basically, 1000ng of total RNA was used for mRNA isolation using NEBNext Ploy(A) mRNA Magnetic Isolation module (New England Biolabs, catalog # E7490 ). Then followed by RNA library construction with NEBNext Ultra II Directional Lib Prep  (New England Biolabs, catalog #E7760) according to the manufacturer's user guide. 12 barcoded libraries were sized on the bioanalyzer, quantitated by QUBIT. Finally, 12 individual libraries were pooled with equimolar and sequenced by Illumina HiSeq 3000 2X100 cycles run for total of 2 runs (Illumina Inc., CA, U.S.)    RNA library construction was performed at the Interdisciplinary Center for Biotechnology Research (ICBR) Gene Expression & Genotyping Core, University of Florida (UF).    HiSeq 3000 sequencing run was performed at the Interdisciplinary Center for Biotechnology Research (ICBR) NextGen DNA Sequencing core, University of Florida (UF).
GEO Accession
GSM3684376