GSM3187659
GSM3187659: HUDEP2_HRI_sgRNA5_undifferentiated_rep2; Homo sapiens; RNA-Seq
RNA samples were harvested in TRIzol™ (ThermoFisher Scientific, Cat #15596018), and purified with the RNeasy Mini kit which included an on-column DNAse treatment to remove genomic DNA (Qiagen, Cat. #74106). Sequencing libraries were then constructed from 100 ng of purified total RNA using the ScriptSeq Complete Kit (Illumina cat# BHMR1224) according to manufacturer's specifications. In brief, the RNA was subjected to rRNA depletion using the Ribo-Zero removal reagents and fragmented. First strand cDNA was synthesized using a 5' tagged random hexamer, and reversely transcribed, followed by annealing of a 5' tagged, 3'-end blocked terminal-tagged oligo for second strand synthesis. The Di-tagged cDNA fragments were purified, barcoded, and PCR-amplified for 15 cycles. The size and quality of each library were then evaluated by Bioanalyzer 2100 (Agilent Techniologies, Santa Clara, CA), and quantified using qPCR.
GEO
GSM3187659
GEO Accession
GSM3187659