GSM3271554: P_8hr_12-1; Homo sapiens; RNA-Seq
Actinomycin D-treated cells were lysed using 1 ml of Trizol (Life Technologies) per 10 cm plate for 5 min at room temperature. RNA was isolated from the lysed cells using the Direct-zol™ RNA MiniPrep Plus kit (Zymo Research) by following the manufacturer's instructions.The concentration of RNA was measured using a Nanodrop 2000c (Thermo Fisher Scientific). cDNA libraries enriched in 3' end fragrments were generated with Nextera kit using Smart-seq2 cDNA amplification method (Picelli et al., PMID: 24385147). Common forward primers were used for all samples; reverse primers with a unique barcode sequence (i5 indices) were specific for each sample. The size distribution of the cDNA library was confirmed on a Bioanalyzer 2100 Instrument (Agilent Technologies). Libraries (150-1000 bp) were subjected to gel size selection to enrich for fragments in the 180 – 280 size range. The concentration of the final library was measured on a qubit fluorometer (Thermo Fisher Scientific).
GEO Accession
GSM3271554