GSM8366271: multiplexed samples (aggregates/blastoids; 0mM/4mM dm-aKG), GEX; Homo sapiens; RNA-Seq
120h single cavity blastoid structures were selected by mouth pipetting. 40h aggregates and 120h blastoids were dissociated into a single cell suspension by incubation with a mixture of TrypLE™ 10x (1x, Thermo Fisher Scientific, A1217701) and Accutase (Merck Sigma Aldrich, A6964) for about 10 to 20 minutes respectively with intermittent pipetting. Each sample was incubated with 0.5 μg of unique hashtag antibody for 20 min on ice. TotalSeq hashtag antibodies (A0251-A0256, Biolegend) were used to multiplex the samples (DOI: 10.1126/sciadv.adg4055). All samples were sorted on a BD FACSymphony S6 (BD Bioscences) resulting in a total cell recovery of around 8000 or 1600 cells, for mRNA2 and mRNA3 respectively, which were loaded onto a Chromium Next GEM chip (10x Genomics). Library preparation was performed according to the Chromium Next GEM Single Cell 3' v3.1 user guide with the addition of the hashtag library for demultiplexing. Combined libraries were sequenced using the NextSeq2000 P2-100 kit (Illumina) with Paired-end sequencing