RNA EXTRACTION/PURIFICATION PROTOCOL FOR TRUCULTURE SAMPLES APP NOTE

Abstract

A continued challenge in investigating human immunity is the ability to correlate in vitro data to in vivo responses. These immune-phenotyping studies are important for basic research to characterize immune activity and function, as well as for pharmacodynamics in drug development to understand drug dosing, safety, and efficacy. While sample analyses have steadily been standardized, the variability of sample collection and culturing remains unresolved. TruCulture® is a whole blood collection and incubation system that reduces and eliminates variables introduced by traditional culturing of peripheral blood mononuclear cells (PBMCs). By culturing whole blood, TruCulture retains all leukocyte populations, all Fc receptor bearing cells, and all circulating soluble factors. Compared to PBMCs, TruCulture more faithfully reflects the in vivo immuneenvironment, allowing more accurate profiling of in vivo responses under stimulating conditions. As TruCulture retains both supernatant and the cell pellet, both gene expression profiling and protein secretion analysis can be performed. This technical note will describe a validated protocol for RNA extraction from TruCulture samples for downstream cDNA analysis or RNAseq.