GSM6701706: THP1-Macrophage, 0hr, Knockdown, Uninfected (R12148); Homo sapiens; RNA-Seq
Total RNA was extracted from THP-1 macrophages, following the double extraction protocol: RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction (TRIzol) followed by a Qiagen RNeasy Micro clean-up procedure. RNA was analyzed on Agilent Bioanalyser for quality assessment with RNA Integrity Number (RIN) range from 7.5 to 10 and median of RIN 9.8. cDNA libraries were prepared using 2 ng of total RNA using the SMARTSeq v2 protocol (Picelli et al., 2014) with the following modifications: 1. Addition of 20 μM TSO; 2. Use of 200 pg cDNA with 1/5 reaction of Illumina Nextera XT kit. The length distribution of the cDNA libraries was monitored using a DNA High Sensitivity Reagent Kit on the Perkin Elmer Labchip. All samples were subjected to an indexed paired-end sequencing run of 2x151 cycles on an Illumina HiSeq 4000 system (28 samples/lane). 2 X 151bp.