GSM9076693: Neutrophils in clonal hemaptopoiesis rhapsody transcriptome; Mus musculus; RNA-Seq
For the generation of single-cell whole-transcriptomes we used a BD Rhapsody system according to manufacture instructions. 60,000 sorted live cells were loaded into a Rhapsody Single Cell Analysis System cartridge. Cell capture and cDNA synthesis were performed according to manufacturer's instructions; cells were isolated into nanowells by gravity, then cells lysed and mRNA, AbSeq oligos and Sample tags oligos were released and captured by beads present in the nanowells. Two separated indexed libraries were prepared for whole transcriptome 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).