GSM4566130: G_DC1_1; Homo sapiens; RNA-Seq
in vitro differentiated cells were FACS sorted in 96-well plates (100 cells/well) containing lysis buffer (RLT). RNA-seq libraries were prepared using Labcyte Echo 525 contactless liquid handling system (Labcyte Inc). In brief, ERCC mix (Thermo Fisher) was added to each sample and first strand full length cDNA was generated with a modified protocol of the SMARTseq v4 Ultra Low Input RNA Kit (Takara Clontech) using poly dT primers and a template switching oligo. Full length cDNA was amplified using SeqAmp™ DNA Polymerase (Takara Clontech). 12 ng of amplified cDNA from each sample was used to generate non-stranded RNA libraries using a modified protocol of the Ovation Ultralow System V2 1-96 kit (NuGEN). In brief, amplified cDNA was fragmented through sonication on Covaris E220 (Covaris Inc), repaired and polished followed by ligation of indexed adapters. Adapter ligated cDNA were pooled before final amplification to add flow cell primers. Libraries were sequenced on HiSeq2500 (Illumina Cambridge) for 100 cycles PE in Rapid mode.
GEO Accession
GSM4566130