Construction of DNA-Origami

Construction Material

Scaffold

1. P8064 Powder and Stock Solution at 150 nM

Staples

• Stock solutions 1 (Hexagonal-prism DNA origami) in three 96‑well plates at 100 uM

1. Schematic Diagram

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

• 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

1. 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

1. Six times ultrafiltration using a 100 kDa molecular-weight-cutoff filter(MWCO)
2. 2% agarose gel electrophoresis running at 90 V for 2 h. Louding(Marker, Scaffold, Samples without ultrafiltration, Samples with ultrafiltration, Marker)
3. Image development

• 3. TME

1. Observation by negative‑stain transmission electron microscopy