A Molecular Logic Gate Based on DNA Origami and Nano-Metal Particle Fluorescent Probes
A technology of nano-metal particles and fluorescent probes, applied in the field of molecular logic gates, can solve the problems of discounting the reliability of experimental results and verifying nano-devices, and achieves the effects of sensitive structural response, reasonable structure, and improved local concentration
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
Method used
Image
Examples
Embodiment 1
[0075] Example 1: AND logic gate
[0076] A kind of AND logic gate based on DNA origami and nano-gold particle fluorescent probe, the construction method comprises the following steps:
[0077] (1) Put 1.6nM M13mp18 chain and staple chain at a molar ratio of 1:10 in 1×TE, 11mM MgCl 2 Mix in the buffer, anneal from 90°C to room temperature, assemble into a dumbbell-shaped DNA Origami, and extend the cleavage strand 1 and cleavage strand 2 on the groove sides at both ends of the DNA Origami "dumbbell";
[0078] (2) Treat the colloidal gold with BSPP overnight, add NaCl until the solution turns purple, centrifuge at 6500rmp for 15min, extract the supernatant, and reclaim it with 1×BSPP, 0.5×TBE, 50mM NaCl solution; mixed with CAP chains in 0.5×TBE, 50mM NaCl solution at a molar ratio of 10:1), and reacted at room temperature for more than 6 hours to obtain gold nanoparticles modified with fluorescent DNA chains.
[0079] (3) Ultracentrifuge the gold nanoparticles modified with ...
Embodiment 2
[0087] Example 2: OR logic gate
[0088] A kind of OR logic gate based on DNA origami and nano-gold particle fluorescent probe, the construction method comprises the following steps:
[0089] Steps (1)-(3) are the same as in Example 1.
[0090] (4) Put the purified DNA Origami and nano-gold particle composite structure sample into fluorescent PCR to collect fluorescent signals. Add 1U / μL protease (Nb.BbvCI for protease 1 and / or Nb.BsrDI for protease 2) during signal acquisition, observe the output of the fluorescent signal, and verify the logical OR gate. like Figure 5As shown, the cleavage sites specifically recognized by the two enzymes are not the same. The two proteases (dark and light) and the corresponding two sets of cleavage sites are shown in different gray levels in the figure, which is logical The construction of the OR gate provides the possibility.
[0091] The sequence of the cleavage chain in this example is shown in Table 5.
[0092] table 5
[0093] ...
Embodiment 3
[0097] Example 3: Three Input Logic Gate
[0098] A three-input logic gate based on DNA origami and nano-gold particle fluorescent probes, the construction method includes the following steps:
[0099] Steps (1)-(3) are the same as in Example 1.
[0100] (4) Put the purified DNA Origami and nano-gold particle composite structure sample into fluorescent PCR to collect fluorescent signals. Add 1U / μL protease (protease is Nb.BbvCI), and / or B chain 4 times the concentration of the cleaved chain, and / or B' chain during the signal acquisition process, observe the output of the fluorescent signal, and verify the three-input logic gate.
[0101] The sequences of the cut chain, B chain and B' chain in this embodiment are shown in Table 7.
[0102] Table 7
[0103]
[0104] In the construction of three-input logic gates, the cutting chain is a "stem-loop" structure, such as Figure 6 shown. In the initial state, the two ends of the cleavage chain have E regions and e regions res...
PUM
Login to View More Abstract
Description
Claims
Application Information
Login to View More