Single-cell sequencing of Pelochelys cantorii
The library synthesis and single cell RNA-sequencing were performed by Gene Denovo (Guangzhou, China). In detail, single hemocytes were barcode-labeled and mixed with reverse transcriptase, and then wrapped by oil surfactant droplets which located in the microfluidic "double cross" system to form Gel Beads-In-Emulsions (GEMs). GEMs flowed into the reservoir and were collected, then silane magnetic beads were used to remove leftover biochemical reagents and primers from the post GEM reaction mixture. Subsequently, full-length, barcoded cDNAs were then amplified by PCR with the sequencing primer R1 and P5 arm, and subsequently sequenced on the Illumina 10x Genomics Chromium platform (Illumina Novaseq 6000).