Team kansai result

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Top Team Project Results Protocols photos

Stage

 First, We constructed the stage and the result is shown in Fig. 5 (a). As you can see, the stage was successfully constructed. But most stages make complexes by blunt-end π–π stacking interactions. To suppress making complexes, thymine spacers were inserted in staple DNAs which is located at side of the stage.

Convex DNA origami

 Next, we tried to construct the convex DNA origami and the result is shown in Fig. 5 (b). Convex DNA origami was successfully constructed.

Stage + Convex DNA origami

 It was confirmed that Stage and Convex DNA origami were constructed. Next, we tried to combine them. The result is shown in Fig. 5 (c) and confirmed that Stage was combined with Convex DNA origami which width was observed ~17 nm. At first time, the yield was ~20% and a reason of the low yield is that the mixture was only incubated at room temperature without slowly annealing. After this, Stage + Convex DNA origami was constructed with slowly annealling from 40℃ to room temperature and the yield rose up to ~60%.

Erectrophoresis

Formation of Stage + Convex DNA origami was confirmed by erectrophoresis and the result is shown in Fig. 6. It was confirmed to construct Stage and Convex DNA origami because of the bands of Lane 2 (in red parentheses) and Lane 3 (in orange parentheses) have different electrophoretic mobility compared with Lane 1.

Molecular NINJA

Fig. 5. AFM images of (a) Stage, (b) Convex DNA origami and (c) Stage + Convex DNA origami.
Fig. 6. The result of electrophoresis. Lane 1, M13mp18 ssDNA. Lane 2, Stage. Lane 3, Convex DNA origami. Lane 4, Stage + Convex DNA origami. (Raw data)