GSM9011918: CTRL_B25; Homo sapiens; RNA-Seq
For RNA extraction from human peripheral blood and kidney tissue samples  intended for 10x Genomics single-cell RNA sequencing, the process typically begins with the dissociation of cells to obtain a single-cell suspension. This is followed by cell lysis, where cells are lysed to release RNA while preserving the integrity of the RNA molecules. The lysate is then treated with a phenol-chloroform extraction or processed using a commercial RNA isolation kit to purify the RNA. Care is taken to minimize RNase contamination throughout the process, ensuring high-quality RNA suitable for downstream 10x Genomics library preparation and sequencing. To prepare a 10x Genomics single-cell RNA-seq library, start by suspending the single-cell sample in a buffer solution. The cell suspension is loaded onto the 10x Genomics Chromium Controller, where individual cells are encapsulated into nanoliter-sized droplets along with barcoded gel beads and reagents. Inside each droplet, cells are lysed, and the mRNA is captured by barcoded oligonucleotides on the gel beads. The barcoded mRNA is then reverse transcribed into cDNA. After breaking the emulsion, the cDNA is purified and amplified using PCR. The resulting cDNA is then fragmented, and sequencing adapters are added to create the final library, which is ready for sequencing on a compatible high-throughput platform.