RT-qPCR (primary cells)

This protocol describes a qPCR-based method for analyzing Atp13a4 expression in primary astrocytes and neurons. Using a 96-well format, reactions are prepared with Fast SYBR Green Master Mix, specific forward and reverse primers, and nuclease-free water. Each sample is analyzed in technical replicates, with no-cDNA controls included to ensure specificity. Ct values are normalized to the housekeeping gene 18S for accurate quantification. Primer sequences and details for Atp13a4 and 18S are provided. This streamlined workflow enables reliable and reproducible assessment of gene expression and is accessible for further customization.