Comparison between healthy individuals and Alzheimer's disease patients with covid
Comparison between healthy individuals and Alzheimer's disease patients with covid
For cell isolation and purification from individuals, peripheral blood mononuclear cells (PBMCs) were collected from severe COVID-19 patients and healthy controls, cryopreserved, and stored in liquid nitrogen. The PBMCs were then thawed, revived, and CD14+ monocytes were isolated using magnetic bead separation. This process involved resuspension in binding buffer, addition of CD14 magnetic beads, incubation, washing, and magnetic separation to collect the CD14+ monocytes For RNA purification, total RNA was extracted from the isolated CD14+ monocytes using the Qiagen RNeasy kit following the manufacturer's instructions. This process included cell lysis, addition of ethanol to adjust binding conditions, loading onto a silica membrane column, washing to remove impurities, and elution of RNA. The quality and quantity of RNA were assessed with a NanoDrop spectrophotometer and Agilent Bioanalyzer, ensuring suitability for uploading to a public repository. RNA was purified from  CD14+ monocytes. PolyA selected strand-specific libraries were generated with 800ug of input RNA using the Illumina Stranded mRNA Prep (Illumina document 1000000124518 v03). Samples were sized, quantified and validated on a Bioanalyzer.