title: GSM8766934: LYZKL; Mus musculus; RNA-Seq
library_name: GSM8766934
library_construction_protocol: the CD4pos and CD8pos population was isolated as described above. Single-cell RNA-seq libraries were generated using Chromium Single cell 3' Library & Gel Bead Kit v3.0 Kits (10×Genomics) according to the manufacturer's protocol. Briefly, about 105 single cells were suspended in calcium- and magnesium-free PBS containing 0.04% weight/volume BSA. About 15000-25000 cells were added to each channel to generate single-cell gel-beads in emulsion (GEMs) for a targeted cell recovery estimate of 10000 single cells (6609 and 7202 for WTKL and LYZKL mice, respectively) After generation of GEMs, reverse transcription reactions were engaged barcoded full-length cDNA followed by the disruption of emulsions using the recovery agent and cDNA clean up with DynaBeads Myone Silane Beads (Thermo Fisher Scientific). The cDNAs were then amplified by PCR with 12 cycles. Subsequently, the amplified cDNA was fragmented, end-repaired, A-tailed, index adaptor ligated and library amplification via PCR (8 cycles). The purified libraries were sequenced on the Illumina NovaSeq 6000 with 28 cycles of read1, 8 cycles of i7 index, and 91 cycles of read2. And finally, 150 bp paired-end reads were generated.
identifiers.namespace: GEO
identifiers.id: GSM8766934_r1