Method development for extraction and purification of dermal RNA from FVB/N mice treated with environmental PAH mixtures

Epidermal RNA samples from FVB/N mice treated with PAH standards and\ud environmental PAH mixtures were harvested and purified using Trizol extraction and\ud RNeasy mini prep kit purification. Epidermal RNA integrity was low (RIN mean 3.5),\ud indicating significant RNA degradation and the need to modify handling, collection, and\ud processing of skin to limit degradation. Potential sources of RNA degrdation include\ud RNase activity, both endogenous and exogenous, thermal degradation during heat-induced\ud epidermal-dermal seperation, and contamination of stock solutions. Testing of\ud the modified method produced dermal RNA samples with acceptable RIN integrity levels\ud (RIN mean 7.6). Variations of the modified method were performed and compared to\ud one another for method optimization