title: GSM8263653: 267608-Blastoids treated with Rapalink-1 for 3 days; Homo sapiens; RNA-Seq
library_construction_protocol: The structures (human blastoids, rapalink-1 treated blastoids and potimplantation cultures dissociated into single cells. Live cells were enriched using EasySep™ Dead Cell Removal (Annexin V) Kit removal kit (Stem cell technologies). Cells counting was performed using nucleocounter NC-250 (ChemoMetec). Desired number of cells were used for generation of Gel Beads in Emulsions (GEMs) using 10X Genomics chromium X at VBCF-NGS, Vienna. For 96 HR samples, 1000 cells were targeted using Chromium Next GEM Single Cell 3ʹ LT Kit v3.1 (PN-1000325) on chip L (PN-1000321) and for post implantation samples 10000 cells were targeted using Chromium Next GEM Single Cell 3' GEM, Library & Gel Bead Kit v3.1 (PN-1000128) on chip G (PN-1000127) according to manufacture's protocols (CG000399 and CG000315, respectively (10X Genomics, Pleasanton, CA)). cDNA were amplified for 11 cycles. Libraries were purified using Ampure XP bead purification protocol and were analyzed using fragment analyzer and qPCR for distributtion and quantity. Libraries were pooled together and sequenced on NovaSeq instrument series flowcell (Illumina, San Diego, CA) using paired end readmode for a total of 300 cycles.