GSM5224811: SH-SY5Y untreated cells, mRNA_library; Homo sapiens; RNA-Seq
Untreated cells were splitted into 2 separate flasks, and left growing to reach about 80% confluence berfore harvesting and sequencing. Cell hashing was performed following manufacturer's instructions (TotalSeq™-A Cell Hashing and TotalSeq™-A Antibodies Protocol for Simultaneous Proteomics and Transcriptomics with 10X Single Cell 3' Reagent Kit v2; BioLegend, San Diego, CA, USA). Briefly, after counting and viability assessment, single cells suspension samples were incubated 10 min on ice with Human TruStain FcX™ Fc Blocking reagent (BioLegend). Next, sample-specific TotalSeq A antibodies (TotalSeq-A0251 anti-human Hashtag 1 Antibody and TotalSeq-A0252 anti-human Hashtag 2 Antibody, BioLegend) were added with subsequent incubation on ice for 30 min. Cells were washed three times with Cell Staining Buffer (BioLegend, Cat# 420201) and finally resuspended in PBS. Cells were counted in an automatic cell counter (Cell counter Thermo Scientific) and samples pooled together. Primers to cDNA PCR (HTO PCR additive primer) were added to increase yield of Hashtag oligonucleotide (HTO, cDNA derived from the TotalSeq™ Hashtag antibodies). After cDNA amplification, HTO-derived cDNAs (180bp) and mRNA-derived cDNAs (>300bp) 2.0 Fluorometer (Invitrogen®, Waltham, MA, USA) and Agilent Bioanalyzer DNA assay (Agilent®, Santa Clara, CA, USA). Libraries were prepared for sequencing following manufacturer's instructions (Illumina®, San Diego, CA, USA) scRNA-seq 10X Genomics
GEO Accession
GSM5224811