GSM4411839: iPSdiff_1_S17; Homo sapiens; RNA-Seq
Day 91 differentiation cultures were dissociated for 30 mins at 37oC using Accutase (Stemcell Technologies, 7920) to obtain single-cell suspensions. Cells were stained with DAPI and sorted for live single cells followed by library preparation. Single-cell RNA-seq libraries were prepared per the Single Cell3'  v2 Reagent Kits User Guide (10x Genomics, Pleasanton, California). Cellular suspensions were loaded on a Chromium Controller instrument (10X Genomics) to generate single-cell Gel Bead-In-EMulsions (GEMs). GEM-RT were performed in a Veriti 96-well thermal cycler (Thermo Fisher Scientific, Waltham, MA). After RT, GEMs were harvested and the cDNAs were amplified, cleaned up with SPRIselect Reagent Kit.  Indexed sequencing libraries were constructed using Chromium Single-Cell 3' Library Kit for enzymatic fragmentation, end-repair, A-tailing, adapter ligation, ligation cleanup, sample index PCR, and PCR cleanup. The barcoded sequencing libraries were quantified by quantitative PCR using the KAPA Library Quantification Kit (KAPA Biosystems, Wilmington, MA). Sequencing libraries were loaded on a NextSeq500 (Illumina, San Diego, CA)  to obtain a sequencing depth of 200K~250K reads per sample.
GEO Accession
GSM4411839