GSM5017829: HUB_DB_004; Danio rerio; RNA-Seq
Live cells were sorted into 384-well plates with Vapor-Lock oil containing a droplet with barcoded primers, spike-in RNA and dNTPs, followed by heat-induced cell lysis and cDNA syntheses using a robotic liquid handler. After cell lysis for 5 minutes at 65C, RT and second strand mixes were distributed with the Nanodrop II liquid handling platform. Primers consisted of a 24bp polyT stretch, a 4bp random molecular barcode (UMI), a cell-specific 8bp barcode, the 5' Illumina Truseq small RNA kit adapter and a T7 promoter. After all cells were pooled in one library, the CEL-seq2 protocol was used for librabry prep (Hashimony et al., 2016)