XIE/Enzyme:2025/Dec

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Revision as of 09:33, 10 December 2025 by Xie guangneng (talk | contribs) (→‎12.09)

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