Team kansai protocols: Difference between revisions
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== <font face="Helvetica" color="#298cda">Materials</font> == | == <font face="Helvetica" color="#298cda">Materials</font> == | ||
Staple DNA strands were purchased from Sigma Genosys (Hokkaido, Japan) and used without further purification. | <font face="Helvetica">Staple DNA strands were purchased from Sigma Genosys (Hokkaido, Japan) and used without further purification.</font> | ||
== <font face="Helvetica" color="#298cda">Self-assembly of DNA origami (Stage, convex DNA origami)</font> == | == <font face="Helvetica" color="#298cda">Self-assembly of DNA origami (Stage, convex DNA origami)</font> == | ||
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. | <font face="Helvetica">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.</font> | ||
== <font face="Helvetica" color="#298cda">AFM observation</font> == | == <font face="Helvetica" color="#298cda">AFM observation</font> == | ||
AFM imaging of DNA origami was performed on a SPA-300HV 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). | <font face="Helvetica">AFM imaging of DNA origami was performed on a SPA-300HV 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).</font> | ||
Revision as of 16:53, 28 September 2011
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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 (Stage, convex 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.
AFM observation
AFM imaging of DNA origami was performed on a SPA-300HV 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).
