Rapid Assembly of DNA Origami in Microfluidic Temperature Gradient

DNA nanostructures, called DNA origami, are self-assembled through DNA hybridization by annealing process. DNA origami consists of a long single-strand DNA, scaffold, and hundreds of complementary oligonucleotides, staple, and constructs various 2D or 3D nanostructures. For DNA origami folding, it is necessary to denature DNAs and annealed them slowly. Although in general annealing process using microtube and commercial thermal cycler, it takes a long time for DNA hybridization due to large scale reactor. Here, We present an effect of temperature distribution during a rapid folding of DNA nanostructures, called DNA origami. DNA origami can fabricate various designs and sizes of 2D/3D nanostructures by self-assembly of DNA hybridization. Based on results of computational fluid dynamics (CFD) simulation, time-dependent temperature distribution in microtube effects the yield of DNA origami. Triangle DNA origami can be folded at -30 °C/min in microfluidic channel whereas no DNA nanostructures were observed by general annealing process. We confirmed 20 times-faster self-assembly of DNA nanostructures in microfluidic channel, compared to general annealing process in microtube by thermal cycler.