Biomod/2014/Kansai/Protocols

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・Materials Staples DNA were purchased from IDT ( U.S.A )

Each staple DNA were dissolved by sterilized water that the concentration of each solution might be 100 μM.

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These solutions were taken 2 μL each and mixed.

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The volume of the solution was diluted with sterilized water to 500μL. we prepared staple DNA mixture.The sample (50 μM) was prepared mixing M13mp18 (4nM), staple DNA mixture (20nM) and 1×TAE/Mg2+ (12.5 mM). This mixture was kept at 90˚C for 10 minutes and cooled from 90˚C to 25˚C at a rate of -1.0˚C/min to anneal the strands.

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The annealed mixture (1μL) was dripped on mica and added 1×TAE/Mg2+ (12.5 mM).

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We observed this product by AFM. the imaging was performed in the fluid Tapping mode with a BL-AC40TS tip (Olympus, Japan) and performed on a Multimode 8/ Nanoscope system (Bruker AVS).

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Self assembly of DNA origami
The formation of DNA origami was performed with M13mp18 ssDNA (4 nM, Takara, Japan), staples DNA (20 nM for each strand) in a solution containing Tris (40 mM), acetic acid (20 mM), EDTA (10 mM), and magnesium acetate (12.5 mM, 1 X TAE/Mg2＋ buffer, 50 µL). This mixture was cooled from 90˚C to 25˚C at a rate of -1.0˚C/min to anneal the strands.

AFM observation
The mixture (1 µL) was deposited on freshly cleaved mica, additional 1X TAE/Mg2＋ buffer (40 µL) was added, and