title: GSM9059462: S3D1 NC1; Homo sapiens; RNA-Seq
library_name: GSM9059462
library_construction_protocol: The harvested stem cell islets were treated with Triple solution for 30 minutes at 37 °C with shaking at 95 rpm. Following treatment, the islets were washed with 1× DPBS and viability was assessed, ensuring a viability rate of over 90%. The cells were then stored in a buffer containing PBS supplemented with 1% w/v BSA. Subsequently, the Single Cell 3.1 protocol was initiated immediately. Library construction was performed according to the manufacturer's instructions (Chromium Single Cell 3' v3.1 protocol, 10x Genomics). Briefly, single cells were resuspended in the master mix and loaded into the Chromium Controller with partitioning oil and barcoded Gel Beads to generate Gel Bead-in-Emulsion (GEMs). Within each GEM, the poly-A RNA from the cell lysate was reverse transcribed into cDNA. This cDNA contained the Illumina R1 primer sequence, a Unique Molecular Identifier (UMI), and the 10x barcode. The pooled barcoded cDNA was cleaned using Silane DynaBeads, then amplified by PCR. The appropriate fragment sizes were selected using SPRIselect reagent to ensure the cDNA was suitable for library construction. During the library construction, the Illumina R2 primer sequence, paired-end constructs with P5 and P7 adapters, and a sample index were added to each fragment for sequencing.