RNA, specifically mRNA, is the intermediary between DNA and protein. By understanding the levels of RNA found in cells, scientists can better comprehend gene expression and regulation. Therefore, the extraction of high-quality RNA is the first and most vital step in performing many molecular techniques found in molecular biology, genetics, biochemistry and microbiological applications. While DNA can survive for extended periods of time and is relatively stable, RNA is typically short-lived and extremely temperature sensitive. Due to RNA’s disposition to degenerate, extracting RNA from tissues requires a method that will isolate purified RNA while also minimizing degradation.
Hard samples such as skin and bone present a significant challenge for the extraction of RNA. Common methods require freezing the tissue in liquid nitrogen or dry ice and then pulverizing with a mortar and pestle. While freezing gives the researcher more control over their disruption conditions, it is typically a very involved and time-consuming process. Bead mill homogenizers such as the Omni Bead Ruptor Elite™ bead mill homogenizer can quickly and efficiently disrupt tough samples for the extraction of RNA. By selecting the most efficient bead media, high-quality RNA can be extracted for various downstream analyses.
Herein, we evaluate the potential for the extraction of RNA from porcine (Sus scrofa) skin on the Omni Bead Ruptor Elite bead mill homogenizer. The extraction efficiency and analyte integrity were evaluated via RT-PCR.
Figure 1: Average BSA recovered across triplicate samples for both the standard polypropylene tube and the low-binding tube. Bars in blue are the concentrations of BSA before bead beating and bars in yellow are concentrations after 20 seconds of bead beating. All values are in ng/μL.