Team kansai protocols2: Difference between revisions
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== <font face="Helvetica" color="#298cda">Bear Trap DNA origami</font> == | == <font face="Helvetica" color="#298cda">Bear Trap DNA origami</font> == | ||
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Constructed Plus shaped DNA origami is added to Bear Trap DNA origami. | |||
Bear Trap DNA origami will catch Plus shaped DNA origami and be transformed rectangular DNA origami. | |||
</font> </p> | </font> </p> | ||
Revision as of 05:33, 24 October 2012
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Materials
Staple DNA strands were purchased from Sigma Genosys (Hokkaido, Japan) and used without further purification.
Self-assembly of DNA origami (Smily, Cyclops, Bear Trap DNA origami)
The formation of DNA origami was performed with M13mp18 ssDNA (4 nM, Takara, Japan), staple strands (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/Mg 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.
Cyclops to Smily DNA origami
Staple strands in center of Smiley DNA origami is added to Cyclops DNA origami solution. Cyclops DNA origami connected by stacking will be inserted staple strands into the center and transformed to Smiley DNA origami.
Bear Trap DNA origami
Constructed Plus shaped DNA origami is added to Bear Trap DNA origami. Bear Trap DNA origami will catch Plus shaped DNA origami and be transformed rectangular DNA origami.
AFM observation
AFM imaging of DNA origami was performed on a E-sweep system (SII, Japan). The mixture (1 µL) was deposited on freshly cleaved mica, additional 1X TAE/Mg buffer (200 µL) was added, and the imaging was performed in the fluid DFM scanning mode with a BL-AC40TS tip (Olympus, Japan).
Agarose gel electrophoresis
The assembled products were loaded into agarose gels (1.5% agarose in 1×TAE/Mg2+ aqueous buffer) and subject to gel electrophoresis at 100 V for one hour. The gel was stained with GelStar.
