For single cell microfluidic qPCR Fluidigm Advanced Development Protocol #41 for single cell gene expression using SsoFast Evagreen Supermix with Low ROX was used. Single cells were collect by FACS into a solution containing 5 μl CellsDirect™ 2× Reaction Mix (Thermo Scientific, 11753100), 1.0 μl Superscript® III RT Platinum® Taq Mix, 2.0 μl 10× primer mix (as for other microfluidic assays, primers at 500 nM), and 2.0 μl nuclease-free water, and frozen at -20 °C until processed.
