title: GSM6514266: PHZ_Day3, rep 2, scRNA-seq; Mus musculus; RNA-Seq
library_construction_protocol: Spleen cells were isolated and resuspended in spleen isolation media (PBS containing 2% FBS, glucose, EDTA). Kit-positive cells were subsequently isolated by positive selection using biotin-conjugated Anti-mouse CD117(Kit) (clone 2B8) antibody followed by incubation with MojoSort streptavidin conjugated magnetic nanobeads. The sorted cells were resuspended with spleen isolation media and transferred to IMDM meida with 1%FBS before loading into the array. Library was performed according to the Seq-well protocol from Gierahn, et al. (2017, Nat Method). Briefly, beads binding with cells were removed and resuspended with reverse transcription master mix to make single strand cDNA. Then the samples were treated with Exonuclease I and doing PCR using dN-SMRT oligo to do the Whole Transcriptome Amplification. After purification with SPRI beads and quality checking by BioAnalyzer, cDNA Tagmentation was done by Nextera XT kit. And the library was constructed by adding paired-end adapter of P5 and N7 sequences and a sample index were added. Each sample was purified and quality checked before sequencing.