Biomod/2012/Titech/Nano-Jugglers/Results: Difference between revisions

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::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)
::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)
==RESULT==


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Revision as of 00:52, 4 October 2012


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} </style> </head> <BODY> <div id="biomodlink"> <<a href="http://openwetware.org/wiki/Biomod">BIOMOD</a>|<a href="http://openwetware.org/wiki/Biomod/2012">2012</a>|Titech Nano-Jugglers </div> <div id="header"> <div id="navigation"> <div id="menu"> <ul> <li><a href="http://openwetware.org/wiki/Biomod/2012/Titech/Nano-Jugglers"><br>Home<br><br></a></li> <li><a href="http://openwetware.org/wiki/Biomod/2012/Titech/Nano-Jugglers/Team/Students"><br>Team<br><br></a></li> <li><a href="http://openwetware.org/wiki/Biomod/2012/Titech/Nano-Jugglers/Project"><br>Project<br><br></a></li> <li><a href="http://openwetware.org/wiki/Biomod/2012/Titech/Nano-Jugglers/Results">Results<br>&<br>Methods</a></font></li> <li class="ach"><a href="http://openwetware.org/wiki/Biomod/2012/Titech/Nano-Jugglers/Achievements"><br>Achievements<br><br></a> <li class="sup"><a href="http://openwetware.org/wiki/Biomod/2012/Titech/Nano-Jugglers/Protocols"><br>Suppl. Info.<br><br></a></li> <li class="none"><a href="http://openwetware.org/wiki/Biomod/2012/Titech/Nano-Jugglers/Acknowledgement"><br>Acknowledgements<br><br></a></li> </ul> </div> </div> </div> </BODY> </html>

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

Experimental results

<html><A href=#TOP title="Vapor deposition"><IMG width="300px" src="http://openwetware.org/images/a/a5/Vapor_deposition.jpg"></A></html> <html><A href=#TOP title="SAM conjugation"><IMG width="300px" src="http://openwetware.org/images/4/48/Thiol_.jpg"></A></html> <html><A href=#TOP title="EDAC conjugation"><IMG width="300px" src="http://openwetware.org/images/3/3d/EDAC_conjugation.jpg"></A></html>
<html><A href=#TOP title="DNA design"><IMG width="300px" src="http://openwetware.org/images/b/b2/DNA_design.jpg"></A></html> <html><A href=#TOP title="半白金　1μの動画"><IMG width="300px" src="http://openwetware.org/images/8/80/Damy.png"></A></html> <html><A href=#TOP title=DNA hybrudization><IMG width="300px"

src="http://openwetware.org/images/f/f1/DNA_hybrudization.jpg"></A></html>

<html><A href=#TOP title="ハイスピードカメラ"><IMG width="300px" src="http://openwetware.org/images/8/80/Damy.png"></A></html> <html><A href=#TOP><IMG width="300px" src="http://openwetware.org/images/e/ed/%E3%82%AB%E3%82%BF%E3%83%A9%E3%83%BC%E3%82%BC.jpg"></A></html> <html><A href=#TOP title="Azobenzene"><IMG width="300px" src="http://openwetware.org/images/e/ea/Azobenzene.jpg"></A></html>

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.

DNA conjugation

File:Body construction.png

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

DNA Design

    We used NUPACK and designed five strands of DNA.

>>see more Method

Observation of platinum hemisphere

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.

Analysis of platinum by High-speed camera

Energy production by using catalase

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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