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Contents 1 Experimental results 1.1 Vapor deposition of Au and Cr on the polystyrene body 1.2 DNA conjugation 1.2.1 SAM conjugation 1.2.2 EDAC conjugation 1.2.3 Observation Conditions 1.3 DNA Design 1.4 Observation of platinum hemisphere 1.5 DNA hybridization in solution of H2O2 1.6 Analysis of platinum by High-speed camera 1.7 Energy production by using catalase 1.8 Dissociation of azobenzene-modified DNA by UV-light irradiation

We have set 3 goals, that is "Energy production for rail-free movement", "Speeding up", and "Direction control".

Experimental results

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Vapor deposition of Au and Cr on the polystyrene body

>>see Method

Image1	Image2	Image3	Image4
40 micron polystyrene beads	40 micron polystyrene beads with completely covered by gold	40 micron silica beads with half vapor deposition of gold	40 micron polystyrene beads with half vapor deposition of gold

From these pictures, we observed that PVD can make hemisphere membrane of gold. By comparing the color of figure1 and figure2, micro-beads completely covered by gold are more black and have a metallic luster.　 From figure3 , it is possible that the size of 40μm beads can deposit gold as hemisphere in that the colors of beads sphere’s limbs are different .Figure 4 shows that 10 micron sized beads can also deposit gold as hemisphere, because metallic luster color and white color are dotted in one sphere.

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DNA conjugation

SAM conjugation     We bond Au with DNA using thiol. >>see more Method

Image1 shows the state of thiol-modified DNA and gold-deposited cover glass reaction. Spot ① and ④ are phosphate and saline buffer. Spot ② and ⑤ are phosphate and saline buffer that DNA was dissolved in. Spot ③ and ⑥ are phosphate and saline buffer that thiol-modified DNA was dissolved. Final concentration of phosphate and saline buffer are the same in these spot. But ①、②　and ③ are added NaCl concentration immediately after 24 hours of incubation. On the other hand ④, ⑤, and ⑥ are intended to raise the concentration of NaCl every 2 hours after 24 hours of incubation for 6hours, then incubate more 16hours. In image2, gold-deposited cover glass is washed with 3 × SSC, then it was blotted on the surface of the water.

    We observe that there were few signs anything after washing in ①,②,④ and ⑤．On the other hand, ③ and ⑥ were left water. This is expected that this spot of gold surface is covered with DNA by thiol conjugation, and became partially hydrophilic. Other spots are still hydrophobic in that there are no conjugations of DNA.

    From this result, we were determined that thiol conjugation is completed.

	Image1 Incubated for 48 hours at a gold plate	Image2 Suck out the water after incubation	Image3 Washed away gold plate surface by 3 × SSC after sucking out the water
image
condition		Hydrophilic layers are stretched in spot ③　and ⑥	Hydrophilic layers　are still stretched in spot ③　and ⑥

EDAC conjugation     We bond polystyrene-beads with DNA using EDAC. >>see more Method

After the protocol, to confirm that amino modified DNA is bind to polystyrene ,we hybridize the DNA with FAM and observe them under blue light by microscope.(Control experiment)

Added Reagent	Polystyren beads DNA which bind with beads Complementary FAM	Polystyren beads DNA which bind with beads	Polystyren beads Complementary FAM
Under Transmitted Light
Under Blue Light
Fluorescence	observed	Not observed	Not observed

※We use polystyrene beads whose 1/4 was covered with Gold and 1/2 was covered with chromium.

From this result, we concluded that amino modified DNA is bind to polystyrene

Observation Conditions

ISO6400

Exposure time（Transmitted Light）１／１００seconds

Exposure time（Blue Light）２seconds

Magnification １０×４０＝４００

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DNA Design

	We used NUPACK and designed five strands of DNA. >>see more Method

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Observation of platinum hemisphere

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DNA hybridization in solution of H2O2

image of DNA hybridization	We used PAGE electrophoresis to ascertain the stability of DNA duplex in thin H₂O₂ solution 1%~5%. >>see more Method

    This image shows the observation results of DNA hybridization in solution of Hydrogen peroxide for 90minutes.From the picture, the hybridized DNA band appear in lane 1,2,3&8. In lane 4,5,6&7 appears single strand band. Lane1&8 are the same, lane 2 & 3 are added hydrogen peroxide solution (1% and 5% for 1h)for 90 minutes. In Lane 5&7 shows the observation results of single strand DNA (ssDNA) in solution of Hydrogen peroxide. In lane 4&6 are the control band of ssDNA.

    Upper white dotted line represents the same position of Lane 1&8 in that they are the same sample. Comparing between 4.5.6&7, these 4 lines appear lower than the white line, and 4-5and 6-7 are few differences. If hydrogen peroxide affects ssDNA, destroy, tear up or denature, line 5 and 7 will appear in the lower position or becomes unclear. Judging from appearances, differences between positive and negative control ware few.Comparing 1,2,3 and 8,these lines are completely appear in the white bar.Differences between these lines are only concentration of hydrogen peroxide. So, we conclude that there is no influence of hydrogen peroxide for DNA hybridizations however DNA is exposed to hydrogen peroxide within 90minutes.So, we could determine that there is no effect of hydrogen peroxide for dsDNA as well as ssDNA.

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Analysis of platinum by High-speed camera

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Energy production by using catalase

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Dissociation of azobenzene-modified DNA by UV-light irradiation

	Azobenzene including DNA can easily dissociate its duplex by irradiating UV-light. We put this switching system in JET body and enabled its control. >>see more Method

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