alias: GSM4413679
title: GSM4413679: PITRM1-WT iPSCs; Homo sapiens; RNA-Seq
library_construction_protocol: RNA libraries were prepared using standard 10X Genomics 3' Single Cell v2 protocol. scRNA-seq libraries were generated using the 10X Chromium Next GEM Single Cell 3' Reagent Kit v3.1 (Cat. 1000128) according to manufacturer's instructions. Approximately 18000 cells per sample were loaded on the Chromium Next GEM Chip G, which was subsequently run on the Chromium Controller (10X Genomics) to partition cells into gel beads-in-emulsion (GEMs). Cell lysis and reverse transcription of poly-adenylated mRNA occurred within the GEMs and resulted in cDNA with GEM-specific barcodes and transcript-specific unique molecular identifiers (UMIs). After breaking the emulsion, cDNA was amplified (11 cycles), enzymatically fragmented, end-repaired, extended with 3' A-overhangs and ligated to adapters. P5 and P7 sequences as well as sample indices (Chromium i7 Multiplex kit) were added during the final PCR amplification step (7 cycles). The fragment size of the final libraries was determined using the Bioanalyzer High Sensitivity DNA Kit (Agilent). Their concentration was determined using the Qubit dsDNA HS Assay Kit (Thermo Fisher Scientific). scRNA libraries were pooled and paired-end sequenced on the Illumina NovaSeq 6000 platform.
identifiers.namespace: GEO
identifiers.id: GSM4413679