title: GSM4873529: PBS; Mus musculus; RNA-Seq
library_construction_protocol: In brief, 500K cells of each sample were sorted and then centrifuged at 300g x 5 minutes, 4 °C. The supernatant was aspirated, and cells were re-suspended in 100 μl of ice cold 1× PBS containing 0.04% BSA. The cells were then counted with trypan blue to make sure high viability above 98%. Cell concentration was adjusted to exactly 700 cells/μl by ice cold 1× PBS containing 0.04% BSA, and cells were kept on ice. Libraries were prepared using the 10X Genomics Chromium Single Cell 3′ v3 Library and Gel Bead Kit. Single-cell suspensions were loaded onto the Chromium Controller to generate 1,000 single-cell gel beads in emulsion per sample. The cDNA of each sample after amplification of 12 cycles was quantified by Qubit and quality-checked by Bioanalyzer to verify the size distribution of cDNA samples and determine indexed PCR amplification of 15 cycles to yield a sufficient and unbiased library for sequencing. After library quality control by Tapestation, indexed library samples were quantified by KAPA library quantification kit (Universal) and pooled with 15ul of 5 nM for each sample. Pooled library sample was sent to Novogen for sequencing. Pair-end sequencing was performed on Illumina® HiSeq platform, with the read length of paired-end 150 bp at each end plus 8-bp i7 index. 10X Genomics Chromium Single Cell RNA-seq
attributes.tag: GEO Accession
attributes.value: GSM4873529