Construction of DNA-Origami: Difference between revisions
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== Construction Material == | == Construction Material == | ||
=== Scaffold === | === Scaffold === | ||
#'''P8064''' '''Powder and Stock Solution at | #'''P8064''' '''Powder and Stock Solution at 150 nM''' | ||
=== Staples === | === Staples === | ||
*'''Stock solutions 1 (Hexagonal-prism DNA origami) in three 96‑well plates at 100 uM''' | |||
#'''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 | |||
*'''Stock solutions 2 (Unmodified hexagonal‑prism DNA origami) in EP tube at 500 nM''' | |||
== Fabrication == | == Fabrication == | ||
*'''''1. Folfing ''''' | |||
{| class="wikitable" | {| class="wikitable" | ||
! Material | ! Material | ||
| Line 16: | Line 18: | ||
! Volume（50 μL） | ! Volume（50 μL） | ||
|- | |- | ||
| Staples ( | | Staples (500 nM) | ||
| 25 μL | | 25 μL | ||
| 12.5 μL | | 12.5 μL | ||
|- | |- | ||
| P8064 ( | | P8064 (150 nM) | ||
| 10 μL | | 10 μL | ||
| 5 μL | | 5 μL | ||
| Line 32: | Line 34: | ||
| 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''''' | |||
#''' 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''' | |||
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