GSM7765270: HeLaS3-IDRGFP-Rep1; Homo sapiens; RNA-Seq
Total RNA was isolated from HeLaS3 cells transfected or not with IDR-GFP constructs from the same experiment as ATAC-seq experiments, using TRIzol® Reagent (Life Technologies) according to supplier's recommendations. Quality and quantity of total mRNA was ensured by Agilent 5400 Fragment Analyzer system mRNA libraries were constructed using poly A enrichment preparation. Messenger RNA was purified from total RNA using poly-T oligo-attached magnetic beads. After fragmentation, the first strand cDNA was synthesized using random hexamer primers followed by the second strand cDNA synthesis. The library was ready after end repair, A-tailing, adapter ligation, size selection, amplification, and purification. The library was checked with Qubit and real-time PCR for quantification and bioanalyzer for size distribution detection. Quantified libraries will be pooled and sequenced on Illumina platforms, according to effective library concentration and data amount