RNA extraction from cells, spheroids, or organoids encapsulated in alginate norbornene using Maxwell® RSC Plant RNA Kit v1

This protocol details the isolation of RNA from cells, spheroids, or organoids encapsulated in alginate norbornene hydrogels using the Maxwell® RSC Plant RNA Kit and automated Maxwell® RSC instrument. The cells, spheroids, or organoids are first released from the hydrogel matrix by alginate lyase digestion, followed by direct processing with kit reagents in the Maxwell® platform. The expected outcome is recovery of RNA with high yield and purity suitable for downstream applications such as qPCR and sequencing. Key optimizations include removal of the hydrogel prior to RNA extraction to reduce contamination from the matrix components, omission of the homogenization step to minimize contamination from residual matrix leftover, and adjustment of reagent volumes within the kit workflow to maximize RNA recovery in low-yield samples.