XIE/Enzyme:2025/Dec

12.08

• HR-DNA Origami（NO） Folding
• Materials

Material	Volume（100 μL）
Staples (500 nM)	25 μL
P8064 (150 nM)	10 μL
10X PBS	10 μL
H2O	55 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.

• Purification (12.09): Reserve a 10 µL aliquot. Add the remaining 90 µL to a 100‑kDa MWCO centrifugal filter unit and dilute to 500 µL with 1× PBS. Centrifuge at 8,000 g for 1.5 min and discard the filtrate . Add UP Water to 500 ul to the filter unit and repeat the centrifugation; perform this wash step three times. After the final wash, adjust the volume in the filter unit to 100 µL with UP Water, invert the filter unit into a fresh collection tube, and centrifuge at 8,000 g for 1.5 min to collect the concentrate.

12.09

• HR-DNA Origami（NO） Folding
• Materials

Material	Volume（100 μL）
Staples (500 nM)	25 μL
P8064 (150 nM)	10 μL
10X PBS	10 μL
H2O	55 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.

• Purification (12.10): Reserve a 10 µL aliquot. Add the remaining 90 µL to a 100‑kDa MWCO centrifugal filter unit and dilute to 500 µL with 1× PBS. Centrifuge at 8,000 g for 1.5 min and discard the filtrate . Add 1X PBS to 500 ul to the filter unit and repeat the centrifugation; perform this wash step three times. After the final wash, adjust the volume in the filter unit to 100 µL with 1X PBS, invert the filter unit into a fresh collection tube, and centrifuge at 8,000 g for 1.5 min to collect the concentrate.

12.10

• Agarose Gel Electrophoresis
• Gel Preparation

1. Recipe: 2.4 g agarose in 120 mL 0.5× TBE
2. Heat in a microwave to dissolve, cool slightly, adjust volume with 0.5× TBE to 120 ul, then add 1.2 mL of 1 M MgCl2 and 12 µL GoldView DNA stain.
3. Insert comb and waite for the gel to cool and solidify.

• Sample preparation

1. Origami (15 nM): sample:loading buffer = 5:1.
2. Scaffold: take 1 µL scaffold, dilute to 10 µL (final concentration 15 nM); scaffold:loading buffer = 5:1.

• Electrophoresis

1. Run at 90 V for 120 min in 0.5× TBE with 10 mM MgCl2
2. Marker、Scaffold（1X）、Origami（10.08-未洗）、Origami（10.08—清洗）、Origami（10.09-未洗）、Origami（10.09-清洗）、Scaffold（10X）、Marker

Origami(NO)