title: GSM4105916: Skin-derived fibroblasts Patient 2; Homo sapiens; RNA-Seq
library_construction_protocol: Dissociation from cell culture dish and resuspension in BSA 1%/HBSS Libraries were prepared according to 10X Genomics ChromiumTM Single Cell 3' Reagent Guidelines v2  for Patient 1 and v3 for Patients 2 and 3 Chemistry as per the manufacturer's protocol. In brief, single cells were sorted into 1% BSA–PBS and partitioned into Gel Bead-In-EMulsions (GEMs) using 10xTM GemCodeTM Technology. This process lysed cells and enabled barcoded reverse transcription of RNA, generating full-length cDNA from poly-adenylated mRNA. DynaBeads® MyOneTM Silane magnetic beads were used to remove leftover biochemical reagents, then cDNA was amplified by PCR over 10 cycles. Quality control size gating was used to select cDNA amplicon size prior to library construction. Read 1 primer sequences were added to cDNA during GEM incubation. P5 primers, P7 primers, i7 sample index, and Read 2 primer sequences were added during library construction. Quality control and cDNA quantification was performed using Agilent High Sensitivity DNA Kit. Sequencing was performed first using Illumina MiSeq SR50 to approximate the number of recovered cells in each sample.
attributes.tag: GEO Accession
attributes.value: GSM4105916