Construction of DNA-Origami: Difference between revisions
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=== Staples === | === Staples === | ||
*'''Stock solutions(Hexagonal-prism DNA origami) in three 96‑well plates at 100 uM''' | *'''Stock solutions 1 (Hexagonal-prism DNA origami) in three 96‑well plates at 100 uM''' | ||
#'''Schematic Diagram''' | #'''Schematic Diagram''' | ||
[[File:Staples.png|thumb|center|400px|Schematic Diagram]] | [[File:Staples.png|thumb|center|400px|Schematic Diagram]] | ||
For detailed information, please refer to the shared "Origami" file on Feishu | For detailed information, please refer to the shared "Origami" file on Feishu | ||
*'''Stock solutions (Unmodified hexagonal‑prism DNA origami) in EP tube at 500 nM''' | *'''Stock solutions 2 (Unmodified hexagonal‑prism DNA origami) in EP tube at 500 nM''' | ||
== Fabrication == | == Fabrication == | ||
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| 27.5 μL | | 27.5 μL | ||
|} | |} | ||
#'''Program folding in PCR thermocycler : The mixed sample was put on a thermal ramp starting with a rapid heat denaturation at 80 °C for 5 min followed by cooling from 80 °C to 60 °C over 20 min, then slow cooling from 60 °C to 24 °C over 14 h.''' | |||
*'''''2. Purifying''''' | *'''''2. Purifying''''' | ||
#''' Six times ultrafiltration using a 100 kDa molecular-weight-cutoff filter(MWCO)''' | #''' Six times ultrafiltration using a 100 kDa molecular-weight-cutoff filter(MWCO)''' | ||
#''' 2% agarose gel electrophoresis running at 90 V for 2 h''' | #''' 2% agarose gel electrophoresis running at 90 V for 2 h.''' Louding(Marker, Scaffold, Samples without ultrafiltration, Samples with ultrafiltration, Marker) | ||
#''' Image development''' | #''' Image development''' | ||
* | |||
*'''''3. TME ''''' | *'''''3. TME ''''' | ||
* | |||
#'''Observation by negative‑stain transmission electron microscopy''' | #'''Observation by negative‑stain transmission electron microscopy''' | ||
Latest revision as of 14:00, 25 November 2025
Construction Material[edit]
Scaffold[edit]
- P8064 Powder and Stock Solution at 150 nM
Staples[edit]
- Stock solutions 1 (Hexagonal-prism DNA origami) in three 96‑well plates at 100 uM
- Schematic Diagram
For detailed information, please refer to the shared "Origami" file on Feishu
- Stock solutions 2 (Unmodified hexagonal‑prism DNA origami) in EP tube at 500 nM
Fabrication[edit]
- 1. Folfing
| Material | Volume（100 μL） | Volume（50 μL） |
|---|---|---|
| Staples (500 nM) | 25 μL | 12.5 μL |
| P8064 (150 nM) | 10 μL | 5 μL |
| 10X PBS | 10 μL | 5 μL |
| H2O | 55 μL | 27.5 μL |
- Program folding in PCR thermocycler : The mixed sample was put on a thermal ramp starting with a rapid heat denaturation at 80 °C for 5 min followed by cooling from 80 °C to 60 °C over 20 min, then slow cooling from 60 °C to 24 °C over 14 h.
- 2. Purifying
- Six times ultrafiltration using a 100 kDa molecular-weight-cutoff filter(MWCO)
- 2% agarose gel electrophoresis running at 90 V for 2 h. Louding(Marker, Scaffold, Samples without ultrafiltration, Samples with ultrafiltration, Marker)
- Image development
- 3. TME
- Observation by negative‑stain transmission electron microscopy