![]() These structures have been used as novel scaffolds for nanopatterning of various nanoparticles ( 17, 18) and quantum dots ( 19), attachment of carbon nanotubes ( 20), immobilization of biomolecules such as proteins ( 21) and viral capsids ( 22), carriers of drugs ( 23), platform for the analysis of single molecular reactions and processes ( 24), and so on. Origami materials can also be lyophilized and stored for future applications ( 16). The formation of DNA origami can also be controlled by using the external signals such as cationic comb-type copolymers/polyvinyl sulphonic acid ( 14) and DNA-binding dendrons/reducing agents or light ( 15). Since the invention of scaffolded origami, various two- (2D) ( 3) and three-dimensional (3D) ( 7–9) DNA origami have been synthesized, and also self-assembled further to create even larger structures ( 10–13). This method has enabled the synthesis of DNA nanostructures with the dimension of ∼100 nm in diameter, whereas the initially prepared non-scaffolded structures were ∼10–20 nm in size ( 4–6). The structural DNA nanotechnology ( 1, 2) has attracted much attention during the past one and a half decades due to the addition of the scaffolded DNA origami method ( 3) to the field. The globular structure of the native and ligated origami was also found to be altered dynamically and progressively upon ethidium bromide intercalation in a concentration-dependent manner. Further, our studies indicated that the ligation of the staple strands influences the globular structure/planarity of the DNA origami, and the origami is more compact when the staples are ligated. Also, the ligation is found to increase the thermal stability of the origami as low as 5☌ to as high as 20☌, depending on the structure. Under the optimized conditions, up to 10 staples ligation with a maximum ligation efficiency of 55% was achieved. Our results indicated that the ligation takes overnight, efficient at 37☌ rather than the usual 16☌ or room temperature, and typically requires much higher concentration of T4 DNA ligase. Here, we report a detailed analysis and optimization of the conditions for the enzymatic ligation of the staple strands in four types of 2D square lattice DNA origami. However, in DNA nanotechnology, the ligation procedures are neither optimized for the DNA origami nor routinely applied to link the nicks in it. ![]() In molecular biology, enzymatic ligation is commonly used to seal the nicks in the duplex DNA. ![]() Most of the DNA nanotubes/tiles and the DNA origami structures melt below 60☌ due to the presence of discontinuities in the phosphate backbone (i.e., nicks) of the staple strands. The low thermal stability of DNA nanostructures is the major drawback in their practical applications. ![]()
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