GSM8655146: Blastocyst; Bos taurus; RNA-Seq
The snRNA-seq libraries from frozen blastoids and day8 blastocysts were prepared using Chromium Nuclei Isolation Kit (10x Genomics,PN-1000493) with minor modifications from the manufacturer's instructions. In brief, frozen blastoids and day8 blastocysts were transferred to pre-chilled sample dissociation tube and were dissociated with pestle within lysis buffer, then the tube was incubated on ice for 7min. Then the dissociated tissue was pipetted onto assembled nuclei isolation column and centrifuged 16,000 x rcf for 20 sec. After being washed with debris removal buffer and wash buffer, the nuclei pellet was resuspended in 50ul resuspension buffer and performed cell counting. Nucleus were loaded into a 10x Genomics Chromium Chip following manufacturer instruction (10x Genomics, Chromium Next GEM Single Cel 3' Reagent Kit v3.1 Dual Index) and sequenced with an Illumina Novaseq 6000 System (Novogene).