Biomod/2011/TeamJapan/Tokyo/Project/Result of the DNA ciliate body

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Results of the DNA ciliate body


  • Following is the result of electrophoresis. Result 1 is DNA ciliate using NHS and EDC. Result 2 is DNA ciliate using EDAC. Meanings of the numbers above results are here.
  1. deoxyribozyme
  2. substrate
  3. negative control (Zn(2+) 0 mM)
  4. positive control (Zn(2+) 10 mM)
  5. no deoxyribozyme beads (Zn(2+) 0 mM)
  6. no deoxyribozyme beads (Zn(2+) 10 mM)
  7. DNA ciliate (Zn(2+) 0 mM)
  8. DNA ciliate (Zn(2+) 10 mM)
  • Three bands were there. From a top, A is the band of deoxyribozyme, B is the band of substrate (didn’t be cleaved), and C is the band of cleaved substrate. If there is cleaved substrate band, it means deoxyribozyme activity is appeared and deoxyribozyme is attached to polystyrene beads successfully.

[Result 1]

  • PAGE of φ200 nm polystyrene beads using NHS and EDC.
Figure1.the result of PAGE of φ200 nm polystyrene beads using NHS and EDC.


  • PAGE of φ1 um polystyrene beads using NHS and EDC.
Figure2.the result of PAGE of φ200 nm polystyrene beads using NHS and EDC.


[Result 2]

  • PAGE of φ200 nm polystyrene beads using EDAC.
Figure1.the result of PAGE of φ200 nm polystyrene beads using NHS and EDC.

Discussions

  • First, we explain this experimentation’s appropriateness for checking DNA ciliate body. To create DNA ciliate body, it was necessary to attach deoxyrobozyme to micrometer-sized polystyrene beads firmly. Furthermore, we had to check deoxyribozyme activity of DNA ciliate body. In this method, these two things could be checked. First, the degree of fixation can be checked. DNA ciliate is removed before loading to polyacrylamide gel, so if deoxyribozyme can’t be attached to polystyrene beads firmly, the leg band is appeared. Second, the deoxyribozyme activity of DNA ciliate body can be confirmed. If there is deoxyribozyme activity of DNA ciliate body, the cleaved substrate band is appeared. Based on the above, this method is appropriate for checking DNA ciliate body.
  • Second, we explain necessity of lanes. Lanes of 1 to 4 are needed for checking deoxyribozyme activity and the positions of each band of DNAs. Lane 1 and lane 2 are control lanes. The band in lane 1 is deoxyribozyme band, and the band of lane 2 is substrate band. Lane 3 and lane 4 are lanes to check deoxyribozyme activity. The solutions in lane 3 doesn’t contain of Zn(2+), the solution in lane 4 contains of Zn(2+). The cleaved band is appeared in lane 4, but doesn’t be appeared in lane 3. This means normal deoxyribozyme isn’t active in no metal ions solutions. Lane 4’s bottom band means position of cleaved substrate.
Lanes of 5 to 8 are needed for checking deoxyribozyme activity of DNA ciliate body. Lane 5 and 6 are lanes for checking to polystyrene beads. If polystyrene beads had deoxyribozyme activity, the cleaved band would be appeared. Lanes of 7 and 8 are needed for checking DNA ciliate body’s deoxyribozyme activity. If DNA ciliate has normal deoxyribozyme activity, the cleaved band is appeared in lane 8 because metal ions are needed for deoxyribozyme activity.

Conclusion

  • We explain the result of DNA ciliate body’s deoxyribozyme activity. In all 3 results, lanes of 5, 6, and 7 aren’t cleaved, on the other hand, lane 8 is cleaved. This shows all three types of DNA ciliate have normal deoxyribozyme activity. In lane of 8, the band of cleaved substrate in result 2 is clearer than in result 1. In conclusion, developing DNA ciliate body is completed, and DNA ciliate by using EDAC is better than by using EDC+NHS.

Reference

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