Transcriptome research has become increasingly popular to monitor global changes in gene expression. In particular, transcriptome changes between diseases versus healthy states can provide valuable insight into understanding immunoresponse mechanisms in disease states. However, the isolation of high-quality mRNA from tough tissues, such as skin, can represent a significant challenge in research. Herein, we compare two methods for mRNA extraction from two murine skin types with a specific focus on mRNA yields and integrity. Our study demonstrates the efficacy of using bead mill technology as the optimal choice of sample preparation in transcriptomic research.
Figure 1. Total mRNA yields and relative purity: Absorbance values at 260 nm and 280 nm were used to determine total mRNA yield and purity following extraction. mRNA yields were on average 4 times higher when bead milling was performed compared to dounce homogenization. Nucleic acid relative purity was comparable between both methods.