Biomod/2013/Todai/Result

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    <li><a href="#A.Cylinder in barrel by DNA origami">A. Cylinder in barrel</a>
 
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        <ul>
 
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          <li>STEP&nbsp;</a>
 
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          <li><a href="#StepA-1">1&nbsp;</a>
 
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          <li><a href="#StepA-4">4&nbsp;</a>
 
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          <li><a href="#StepA-5">5</a>
 
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Revision as of 10:14, 31 August 2013


Result-Todai nanORFEVRE-

 Result


We focus on pilot study now, so please visit Pilot Study ,too.


 A. Cylinder in barrel by DNA origami

 STEP1. DNA strands assemble to form designed structures.

[Method]

First, the DNA nanostructure, "Cylinder in barrel" , was used for pilot study.

To know the optimum condition of the structure assembly, we did experiments in three conditions as follows.

  • At different concentration of MgCl2
  • At different incubate temperature
  • At different length of incubate time
( protocols )



[Result & Discussion]
To decide optimum concentration of MgCl2
Agarose-gel electrophoresis to research the optimum concentration of MgCl2

Fast-migrating species upon agarose-gel electrophoresis was yielded at 20mM MgCl2 condition. Correctly folded structures were yielded at 20mM of MgCl2, because they were more compact than misfolded versions. (The structures at high concentration (30 and 40mM) of MgCl2 tend to aggregate because of positive charge.)

--->> Optimum concentration of MgCl2: 20mM


To decide optimum incubate temparature
Agarose-gel electrophoresis to research the optimum temperature

Fast-migrating species upon agarose-gel electrophoresis was yielded at 47.7 or 51.6 degree C. The band at 47.7 degree C is more clear than that of at 51.6 degree C. The band of 47.7 degree C is fast migrated and strong.

--->> Optimum temparature : 47.7 degree C


To decide optimum length of incubate time
Agarose-gel electrophoresis to research the optimum time

The structure assembled for 4h becomes most compact. The band for 4h is fast migrated and strong.

--->> Optimum incubate time : 4h











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