Most shapes are assembled in a one-pot reaction and annealed for 90 minutes from 90°C to 20°C uniformally with 10nM M13mp18 and 100nM staple concetration (10:1 ration) with 10mM Magnesium buffer. See shape-specific annealing protocol here. For single-origami assembly we recommend adding extension cover staples for higher yield rate and reduced aggregation.

To start experimental validation of nanoframes, it is generally a good idea to perform gel electrophoresis first, to confirm structure complexity and verify that aggregation is not happening. The example figure below demonstrates the results of gel electrophoresis performed on two identical samples of a 50nm x 50nm x 50nm x 4 cube with extension covers.

Once the strucutral complexity is verified with gel, TEM and AFM follows. We found that using non gel-purified samples gave better readouts on TEM and AFM. TEM samples were washed in water droplets and stained with 2% uranyl acetate for 2 minutes.