H1-CFS EU differentiation protocol
Single cell encapsulation and cDNA libraries were prepared by Chromium Next GEM Single Cell 3 Reagent Kit v3.1 and Chromium Next GEM Chip G Single Cell Kit. Cells in suspension are counted and loaded into individual wells of 10X Chromium Single Cell chip. Single cells are then encapsulated into Gel Beads-in-emulsion (GEM) by 10X Chromium Single Cell Controller. Single Cell 3 Reagent Kit v3.1 is used to perform downstream steps, in brief as follows: reverse transcription is performed on GEMs, followed by cDNA cleanup and amplification. The double-stranded cDNA will then go through enzymatic fragmentation, adapter ligation, index PCR and SPRIselect size selection per manufacturers protocol. Library size and concentration are determined by Qubit, quantitative PCR and Bioanalyzer assays.