Biomod/2014/Attachement: Difference between revisions

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<h1> Team Nanoscooter Braunschweig </h1>

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Particle attachment Binding Nanoparticles to the DNA-Origami To bind the Pt-nanoparticles to the DNA-origami 4 µL of the purified DNA-origami were mixed with 6 µL of the functionalized Pt-nanoparticles and incubated overnight. At the next day the solution was filtered to remove the excess of Pt-nanoparticles. 10 µL of the DNA-origami solution were mixed with 400 µL of the buffer solution (PBS + 100 mM NaCl and 12 mM MgCl2) and centrifuged this at 5 krcf at 4°C 5 minutes in a 100K Amicon® filter. To recover the DNA-origami from the filter, the filter was turned upside down and centrifuged for 3 min at 1 krcf.

Gel electrophoresis To proof the attachment of the platinum particles to the Nanoscooter gel electrophoresis is used. We used a 0.7 % agarose gel and checked for differences between Pt-particles bound to the Nanoscooter and the Pt-particles alone. For the gel, 0.35 g of agarose were added to 50 mL 0.5x TBE buffer and heated for 2 min in the microwave oven. After cooling down 2 µL Sybr® Safe DNA Gel Stain and 800 µL 16 mM aqueous MgCl2 solution were added to the agarose gel. Each sample is mixed with 3 µL 10x BlueJuiceTM Gel Loading buffer. Samples: 1. 10 µL DNA origami + 10 µL functionalized platinum particles + 3 µL 10X BlueJuiceTM Gel Loading buffer 2. 10 µL functionalized platinum particles + 10 µL TE buffer + 11 mM MgCl2 + 3 µL 10X BlueJuiceTM Gel Loading buffer The gel electrophoresis was carried out for 90 min and a potential of 85 V. After that the gel was analyzed under UV radiation.

<div align="center">Figure 1: Gel electrophoresis of the Nanoscooter + platinum particles (slot 1) and the platinum particles (slot 2).</div>

As it can be seen in figure 1 the band of the Nanoscooter with platinum particles (slot 1) runs less far than the platinum particles itself (slot 2). This proves that the attachment of the nanoparticles to the Nanoscooter was successful.

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@@ Line 75: / Line 75: @@
 <font size="+2pt"><u>Particle attachment</u></font> <br><br>
 <font size="+1pt">Binding Nanoparticles to the DNA-Origami</font> <br>
-<p align="justify">To bind the Pt-nanoparticles to the DNA-origami 4 µL of the purified DNA-origami were mixed with 6 µL of the functionalized Pt-nanoparticles and incubated overnight. <br>
+<font size="3pt"><p align="justify">To bind the Pt-nanoparticles to the DNA-origami 4 µL of the purified DNA-origami were mixed with 6 µL of the functionalized Pt-nanoparticles and incubated overnight. <br>
-At the next day the solution was filtered to remove the excess of Pt-nanoparticles. 10 µL of the DNA-origami solution were mixed with 400 µL of the buffer solution (PBS + 100 mM NaCl and 12 mM MgCl2) and centrifuged this at 5 krcf at 4°C 5 minutes in a 100K Amicon® filter. To recover the DNA-origami from the filter, the filter was turned upside down and centrifuged for 3 min at 1 krcf.</p><br><br>
+At the next day the solution was filtered to remove the excess of Pt-nanoparticles. 10 µL of the DNA-origami solution were mixed with 400 µL of the buffer solution (PBS + 100 mM NaCl and 12 mM MgCl<sub>2</sub>) and centrifuged this at 5 krcf at 4°C 5 minutes in a 100K Amicon<sup>®</sup> filter. To recover the DNA-origami from the filter, the filter was turned upside down and centrifuged for 3 min at 1 krcf.</p></font><br><br>
 <font size="+1pt">Gel electrophoresis</font><br>
-<p align="justify">To proof the attachment of the platinum particles to the Nanoscooter gel electrophoresis is used. We used a 0.7 % agarose gel and checked for differences between Pt-particles bound to the Nanoscooter and the Pt-particles alone.<br>
+<font size="3pt"><p align="justify">To proof the attachment of the platinum particles to the Nanoscooter gel electrophoresis is used. <br> We used a 0.7 % agarose gel and checked for differences between Pt-particles bound to the Nanoscooter and the Pt-particles alone. For the gel, 0.35 g of agarose were added to 50 mL 0.5x TBE buffer and heated for 2 min in the microwave oven. After cooling down 2 µL Sybr<sup>®</sup> Safe DNA Gel Stain and 800 µL 16 mM aqueous MgCl<sub>2</sub> solution were added to the agarose gel. Each sample is mixed with 3 µL 10x BlueJuice<sup>TM</sup> Gel Loading buffer.</p><br>
-For the gel, 0.35 g of agarose were added to 50 mL 0.5x TBE buffer and heated for 2 min in the microwave oven. After cooling down 2 µL Sybr® Safe DNA Gel Stain and 800 µL 16 mM aqueous MgCl2 solution were added to the agarose gel. Each sample is mixed with 3 µL 10x BlueJuiceTM Gel Loading buffer.</p><br>
+Samples:<br> 	1. 10 µL DNA origami + 10 µL functionalized platinum particles + 3 µL 10X BlueJuice<sup>TM</sup> Gel Loading buffer<br>
-Samples:<br> 	1. 10 µL DNA origami + 10 µL functionalized platinum particles + 3 µL 10X BlueJuiceTM Gel Loading buffer<br>
+. 10 µL functionalized platinum particles + 10 µL TE buffer + 11 mM MgCl<sub>2</sub> + 3 µL 10X BlueJuice<sup>TM</sup> Gel              Loading buffer<br>
-. 10 µL functionalized platinum particles + 10 µL TE buffer + 11 mM MgCl2 + 3 µL 10X BlueJuiceTM Gel Loading buffer<br>
+<p align="justify">The gel electrophoresis was carried out for 90 min and a potential of 85 V. After that the gel was analyzed under UV radiation. <font size="3pt"></p><br>
-<p align="justify">The gel electrophoresis was carried out for 90 min and a potential of 85 V. After that the gel was analyzed under UV radiation. </font></p><br>
 <div align="center"><img src="http://openwetware.org/images/5/5a/Particle-Attachement-Gel.png" width="" height="" ></div>
-<font size="-2 pt"><div align="center">Figure 1: Gel electrophoresis of the Nanoscooter + platinum particles (slot 1) and the platinum particles (slot 2).</font></div>
+<i><font size="3 pt"><div align="center">Figure 1: Gel electrophoresis of the Nanoscooter + platinum particles (slot 1) and the platinum particles (slot 2).</font></div><br></i>
-<p align="justify">As it can be seen in figure 1 the band of the Nanoscooter with platinum particles (slot 1) runs less far than the platinum particles itself (slot 2). This proves that the attachment of the nanoparticles to the Nanoscooter was successful.</p>
+<font size="3pt"><p align="justify">As it can be seen in figure 1 the band of the Nanoscooter with platinum particles (slot 1) runs less far than the platinum particles itself (slot 2). This proves that the attachment of the nanoparticles to the Nanoscooter was successful.</p></font>
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Biomod/2014/Attachement: Difference between revisions

Revision as of 08:20, 22 October 2014

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