GSM7988956: ESC3; Homo sapiens; RNA-Seq
Total RNA was extracted using a SMART-Seq V4 Ultra-Low Input RNA Sequencing kit (Clontech, CA, USA) that uses oligo(dT) priming. cDNA was synthesized with 3′SMART-Seq CDS primer II (Clontech, CA, USA) and PCR amplified (17 cycles) from 10 pg of RNA following manufacturer's instructions. Amplified cDNA was purified using AMPure XP magnetic beads (Illumina, CA, USA). cDNA integrity was checked with a 2100 Bioanalyzer (Agilent Technologies, CA, USA), then 1 ng of cDNA per sample was fragmented, and libraries were constructed using NexteraXT DNA sample preparation (Illumina, CA, USA) according to manufacturer's instructions. Samples were quantified using a 2100 Bioanalyzer (Agilent Technologies, CA, USA). An RNA pool was generated using equal concentration (5 nM) of RNA per sample. Barcodes and adapters were included in the library to sequence and identify all samples.