Single-cell RNA-seq analysis of Drosophila neuroblast tumors

Inactivation of prospero in Drosophila neuroblasts during larval stages induces unlimited neuroblast amplification leading to tumors that persist growing in adults. Tumors present in the ventral nerve cord of adult flies were dissected, dissociated and GFP-labelled tumor neuroblasts were isolated by FACS. 10000 neuroblasts were then processed for single-cell RNA-seq analysis. Single Cell RNA sequencing library were generated using the 10x Genomics Chromium Platform and sequenced on the Illumina Nextseq 500. eLife 2019;8:e50375 DOI: 10.7554/eLife.50375 FACS sorted tumor neuroblasts were processed using the Chromium Single Cell Controller (10X genomics, Pleasanton). Libraries were generated and sequenced from the cDNA and the 10x Barcodes are used to associate individual reads back to the individual partitions. Analysis using molecular indexing information provides an absolute digital measurement of gene expression levels. Sequencing was performed using a NextSeq 500 Illumina device (1sample) containing transcript lengh of 57 bp.Droplet Based