XIE/3:2026/Jan

1.22

• Origami folding

折叠三种浓度的Origami（50nM,100nM,150nM）

50 nM

Samples	Volume
Scaffolld (1uM)	5 ul
Staples (10uM)	2.5 ul X 9
PBS (10X)	10 ul
Water	62.5 ul

100 nM

Samples	Volume
Scaffolld (1uM)	10 ul
Staples (10uM)	5 ul X 9
PBS (10X)	10 ul
Water	35 ul

150 nM

Samples	Volume
Scaffolld (1uM)	15 ul
Staples (10uM)	7.5 ul X 9
PBS (10X)	10 ul
Water	7.5 ul

• 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 :The incubated samples were mixed with 400 μL of phosphate‑buffered saline (PBS) and transferred into 50 kDa molecular‑weight‑cut‑off (MWCO) centrifugal ultrafiltration units. Samples were centrifuged at 10,000 × g for 3 min, the retentate was brought back to volume with PBS, and the centrifugal ultrafiltration step was repeated; this wash/centrifugation cycle was performed a total of three times

琼脂糖凝胶电泳(1.23)

1. 2.4g琼脂糖加入到120 ml（0.5X TBE）中，微波炉加入至完全溶解；用 0.5X TBE 补足，再加入 1.2ml 1M MgCl2 和 12 ul Goldview；倒入模具，插上梳子。
2. 取Scaffold、Origami（50nM）、Origami（100nM）、Origami（150nM）各 10 ul， 各加入 Loudingbuffer 2 ul，涡旋混匀。
3. Scaffold、Origami（50nM）、Origami（100nM）、Origami（150nM）依次上样，90V 3h（电泳液：0.5X TBE + 10mM MgCl2）

结论：150 nM 浓度太低

1.25

• Origami folding

折叠三种浓度的Origami（100nM,190nM,300nM）

• 300 nM 的Origami是通过超滤浓缩获得

190 nM

Samples	Volume
Scaffolld (10uM)	1.9 ul
Staples (10uM)	9.5 ul X 9
PBS (10X)	10 ul
Water	2.6 ul

100 nM X 4

Samples	Volume
Scaffolld (1uM)	10 ul
Staples (10uM)	5 ul X 9
PBS (10X)	10 ul
Water	35 ul

• 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.