Computational protocol: Folding complex DNA nanostructures from limited sets of reusable sequences

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Protocol publication

[…] Agarose gel-based yield estimation was carried out by using ImageJ (http://rsb.info.nih.gov/ij/). The percentage of structure that ran as a monomeric, leading band was estimated as the background-subtracted integrated intensity value divided by the background-subtracted integrated intensity value enclosing the material from the well, down to the bottom of the leading band. [...] Six microliters of the purified folding reaction product was applied on glow-discharged, carbon-coated, 400 mesh formvar grids (Electron Microscopy Sciences), incubated for 1 min, blotted off and stained with 2% (w/v) aqueous uranyl formate solution. The electron micrographs were collected with a FEI TECNAI T20 transmission electron microscope and a Tietz TVIPS 8k camera at normal magnification of 46,000×. Particles for class averaging were picked and calculated with EMAN 2. The number of particles picked for the class averages in Figure , and  was between 220 and 295. […]

Pipeline specifications

Software tools ImageJ, EMAN
Applications cryo-EM, Microscopic phenotype analysis