GSM8808565: Bulk RNA-seq: SMA Infant #4, Post-Treatment; Homo sapiens; RNA-Seq
Bulk RNA-seq: RNA was extracted from whole blood using a Quick-RNA Whole Blood kit (Cat #R1201, Zymo Research) following the manufacturer's protocol. The quality and integrity of the extracted RNA were assessed using an Agilent Bioanalyzer to ensure sufficient RNA quality for sequencing. Library preparation was then performed using the Illumina TruSeq RNA Sample Preparation Kit, converting the RNA into cDNA for sequencing. Single-cell RNA-seq: All samples were collected in lavender blood collection tubes, transported to the lab on ice, and cryopreserved using the SENSE protocol. PBMCs were isolated using an EasySep human CD15 positive selection kit (Cat #18651, STEMCELL), and viability was confirmed to exceed 80% with the TC20 Automated Cell Counter (Cat #1450102, Bio-Rad). Bulk RNA-seq: The raw sequencing libraries were sequenced on an Illumina NovaSeq 6000 platform (Novogene, Sacramento, CA), generating paired-end 150 bp reads. Single-cell RNA-seq: Single-cell RNA sequencing (scRNA-seq) was performed on viably thawed samples using the Drop-seq approach, which encapsulates single cells and uniquely barcoded primer beads in oil droplets for high-throughput transcriptome analysis. Single-cell suspensions were processed with the 10x Genomics workflow, generating digitally barcoded droplets for cell lysis and library preparation using a 10x Chromium Controller. Libraries were prepared with the 10x Genomics Single Cell 5' v2 and Chromium Next GEM Single Cell 5' v2 reagent kits. Sequencing was conducted on the NovaSeq 6000 platform, achieving an average detection of ~1,000–2,000 genes per cell.