Cell 3′ Gel Beads and then a stream of oil. This ideally encapsulates a single cell and an individual gel bead in an oil suspension. Upon cell lysis and dissolution of the Single Cell 3′ Gel Bead within the droplet, primers containing an Illumina P7 and R2 sequence, a 14 bp 10×Barcode, a 10 bp randomer, and a poly-dT primer sequence were released and mixed with the cell lysate and bead-derived Master Mix. Barcoded, full-length cDNA from poly-adenylated mRNA was then generated in each individual bead. After this, the individual droplets were broken and homogenized before the remaining non-cDNA components were removed with silane magnetic beads. The libraries were then size-selected, and the R2, P5, and P7 sequences were added to each selected cDNA during end repair and adaptor ligation. After Illumina bridge amplification of the cDNA, each library was sequenced using the Illumina HiSeq4000 2×150bp in Rapid Run Mode.
