RNA  nanotechnology  is  gaining  in  popularity  due  to  its simplistic and rational design for creating nanostructures for applications in medicine and therapeutics.  RNA nanosquares were previously designed and constructed demonstrating the tunable size and stability of RNA as a nanoparticle delivery system. Dynamic light scattering (DLS) and atomic force microscopy (AFM) were utilized to characterize the size and shape of the squares verifying the molecular model and native PAGE assembly data.  Here, we display the use of analytical ultracentrifugation  (AUC)  to  further  validate  the  size  of the nanostructures, promoting the use of the instrument as  an  orthogonal  tool  for  nanoparticle  characterization and provide insights into applications where AUC provides distinct  advantages  for  nanoparticle  characterization  over other sizing methods.

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