title: GSM6012657: YO2_counts; Mus musculus; RNA-Seq
library_construction_protocol: Up to 60.000 sorted mouse neutrophils from different tissues were  labelled with BD® AbSeq Ab-Oligos and Sample Tags. Cells were loaded into a Rhapsody Single Cell Analysis System cartridge. Cell capture and cDNA synthesis were performed according to manufacturer's instructions. Basically, cells are isolated into nanowells by gravity, then cells are lysed and mRNA, AbSeq oligos and Sample tags oligos are released and captured by beads present in the nanowells. Each bead contains a unique oligo named “cell label” to identify each individual bead. All beads present in the cartridge are collected and cDNA synthesis takes place in a single reaction. At this point each cDNA, AB-Seq oligo and Sample Tag oligo is attached to its corresponding “cell label” oligo. Three separated indexed libraries were prepared for whole transcriptome analysis, Ab-Seq analysis and Sample Tag demultiplexing following manufacturer's instructions. The average size of the libraries was calculated using the 2100 Bioanalyzer (Agilent) and the concentration was determined using the Qubit® fluorometer (Thermofisher). Finally, libraries were combined and sequenced together in a paired end run (60x42) using a NextSeq 2000 system (Illumina) and a P2 flowcell. Output files were processed with NextSeq 1000/2000 Control Software Suite v1.4.1. FastQ files for each sample were obtained using BCL Convert v3.6.3 software (Illumina).