parental 100 starting cells
The single-cell suspension, bead suspension, and the droplet generation oil were loaded into their respective containers and connected via high precision pumps to the scRNA chip as part of the muEncapsulation system. The flow rates were adjusted so that each droplet generated encapsulated one bead and one cell and monitored under the systems high speed digital microscope. Droplets were collected, and a small aliquot was examined microscopically to ensure uniformity of bead size and occupancy. The beads were washed and used for reverse transcription/cDNA synthesis and PCR according to the drop-seq workflow. The PCR products were purified, pooled, and quantified in a BioAnalyzer High Sensitivity Chip 204 (Agilent). Sequencing libraries were prepared using the Nextera XT DNA sample prep kit (Illumina Inc) following manufacturers protocol. The libraries were purified, quantified, and sequenced by GenWiz on an Illumina High-Seq instrument using 2x150 nt reads.