GSM4977189: CAGE DE in vitro induced definitive endoderm 66h rep3; Homo sapiens; RNA-Seq
Total RNA was extracte using NucleoSpin® RNA ((Macherey-Nagel) 3 μg of total RNA from each sample were subjected to reverse transcription with random primers. The 5-end cap structure was biotinylated and captured with streptavidin-coated magnetic beads (Thermo Fisher). After ligation of 5′ and 3′ adaptors, second-strand cDNA was synthesized, followed by exonuclease I (New England BioLabs) digestion. The purified CAGE libraries were sequenced using single-end reads of 50 bp on the Illumina HiSeq 2500 (Illumina, USA).