DNA tetrahedral nano-structure signal probe and application thereof
A technology of signaling probes and nanostructures, applied in the biological field, can solve the problems of low efficiency of enzyme reactions
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Embodiment 1
[0081] Four single-stranded DNAs used to assemble tetrahedral DNA nanostructure signal probes: tetra-a-reporter (77bp, molecular weight 23718.43, ssDNA); tetra-b-biotin (55bp, molecular weight 17032.04, 5' end modified biotin ssDNA ); tetra-c-biotin (55bp, molecular weight 16909.915' end modified biotin ssDNA); tetra-d-biotin (55bp, molecular weight 16888.84, 5' end modified biotin); all were synthesized by Shanghai Shenggong Company.
[0082] The four DNAs that constitute the tetrahedral nanostructure signal probe contain three structural domains, each of which is complementary to the corresponding structural domains of the other three single-stranded DNAs (17 pairs of bases are complementary), and each single-stranded DNA surrounds the four sides One face of the tetrahedron is a circle, and each vertex contains two bases (non-complementary, flexible) for bending, and the 3' and 5' ends of the single-stranded DNA converge on the four vertices of the tetrahedron. Tetra-a-repor...
Embodiment 2
[0097] The electrochemical detection method is as follows:
[0098] 1) Self-assembled tetrahedral DNA capture probe and tetrahedral DNA nanostructure signal probe
[0099] Synthesis of capture probes: take equal amounts of four single-stranded DNAs of tetra-a, b, c, and d, and use TM buffer (20mM tris, 50mM MgCl 2 ,pH8.0) diluted to a concentration of 1uM and a volume of 100uL. After 95 degrees Celsius for 3 minutes, immediately cool down to 4 degrees Celsius.
[0100] Synthesis of signal probes: take equal amounts of four single-stranded DNAs of tetra-a-reporter, b-biotin, c-biotin, and d-biotin, and use TM buffer (20mM tris, 50mM MgCl 2 ,pH8.0) diluted to a concentration of 1uM and a volume of 100uL. After 95 degrees Celsius for 3 minutes, immediately cool down to 4 degrees Celsius.
[0101] 2) Clean and polish the electrodes and assemble
[0102] Take a gold electrode with a diameter of 2mm, first polish it with 0.3um and 0.5um alumina powder in sequence, then use etha...
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