GSM6410775: E50, embryo B, part 1, scRNAseq; Callithrix jacchus; RNA-Seq
Approximately 1/3 of posterior part of CS12 fetus (Fig XX) were dissociated into single cells with 0.1% Trypsin/EDTA treatment for 15 min at 37 °C with periodic pipetting. After the reaction was quenched by adding an equal volume of FBS strained through a 70 μm nylon cell strainer, cells were resuspended in FACS buffer (0.1% BSA in PBS). Cells were loaded into chromium microfluidic chips with the Chromium Next GEM Single Cell 3ʼReagent Kit (v3.1 chemistry) and then used to generate single-cell gel bead emulsions (GEMs) using the Chromium Controller (10× Genomics) according to the manufacturer's protocol. GEM-RT was performed in a C1000 Touch Thermal Cycler with 96-Deep Well Reaction Module (Bio-Rad). All subsequent cDNA amplification and library construction steps were performed according to the manufacturer's protocol. Libraries were sequenced using a 2 × 150 paired-end sequencing protocol on an Illumina HiSeq 4000 or NovaSeq 6000 instrument.