Chromium single cell b chip

Single‐cell RNA sequencing libraries were prepared using the Chromium Single Cell 3′ Library Gel Bead Kit v3 according to the manufacturer's instructions (10× Genomics). Cryo‐preserved cells were washed and re‐suspended in 0.1% BSA in PBS immediately before loading on a Chromium Single Cell B Chip (10× Genomics), with a target capture of 10,000 cells. Libraries' quality was assessed using the Agilent BioAnalyzer High‐Sensitivity DNA assay then sequenced using Illumina's NextSeq 500 platform.

The scRNA-seq library preparation and sequencing of pregnant mouse uterus samples was performed at the Centre of PanorOmic Science, at the University of Hong Kong. A total of 22,750 cells in each sample were input into the Chromium Single Cell B Chip (#1000073, 10× Genomics), with the aim being to recover 10,000 cells for sequencing. Single cells were encapsulated using the Gel Bead in emulsion using the 10× Chromium Controller. Reverse transcription was performed on the Gel Bead in emulsion [26 (link)] for cDNA synthesis. The cDNA clean-up and amplification were performed using the Chromium Single Cell 30 Library and Gel Bead Kit v3 (#1000075, 10× Genomics). The libraries were sequenced on a NovaSeq 6000 (Illumina, San Diego, CA, USA) using a 151-bp paired-end mode at a depth of approximately 500 million reads per sample.