Gene detection probe and preparation method thereof
A gene detection and probe technology, applied in the field of gene detection probes and their preparation, can solve the problems of single detection method, biotin-labeled fluorescence quenching, detection methods without integrated detection methods, etc. The ability to distinguish between base mismatch sequences and the effect of reducing the influence of detection results
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Embodiment 1
[0026] A gene detection probe comprising a gene detection probe containing biotin, metal Au ions, and radioactive DATP.
[0027] Gene detection probes include the following preparation methods:
[0028] S1, at 100 ° C, AUNPS is prepared by Turkevich-Frens method by Turkevich-Frens method by adding reducing agent and the trival compound in a ratio of mass 100: 1, and a good dispersibility.
[0029] S2, the prepared AUNPS will be mixed by mixing the mixed solution by mixing the mixed solution to the organic solvent, and the mixed solution is treated with centrifugation, and the upper liquid is removed, and then the precipitate is left after drying and obtains the marking nanoparticle.
[0030] S3, by incorporating 1 μg of Template DNA, 2.5 μL of 10 × EcoRI buffer, 1-2u EcoRi enzyme and 25 μl of deionized water, and retained 36.5 ° C for water bath.
[0031] S4, after 2 h 2 h after 36.5 ° C, the water temperature was adjusted to 75 ° C, and 1 μl of nanoparticles, 1 μl of unlabeled DN...
Embodiment 2
[0043] S1, at 100 ° C, AUNPS is prepared by Turkevich-Frens method by Turkevich-Frens method by adding reducing agent and the trival compound in a ratio of mass 100: 1, and a good dispersibility.
[0044] S2, the prepared AUNPS will be mixed by mixing the mixed solution by mixing the mixed solution to the organic solvent, and the mixed solution is treated with centrifugation, and the upper liquid is removed, and then the precipitate is left after drying and obtains the marking nanoparticle.
[0045] S3, by incorporating 1 μg of Template DNA, 2.5 μL of 10 × EcoRI buffer, 1-2u EcoRi enzyme and 25 μl of deionized water, and retained 36.5 ° C for water bath.
[0046] S4, after 2 hours of a water bath at 36.5 ° C, the water temperature was adjusted to 75 ° C, and 1 μl of the Kjeno-enzyme of 1 μl of nanoparticles and 1u was added 15 min to terminate.
[0047] S5, then add 1 g of Tris balanced phenol and chloroform-isoprene 24: 1, severe oscillate for 10 s, 10000 r / min for 10 min, and ...
Embodiment 3
[0053] S3, by incorporating 1 μg of Template DNA, 2.5 μL of 10 × EcoRI buffer, 1-2u EcoRi enzyme and 25 μl of deionized water, and retained 36.5 ° C for water bath.
[0054] S4, after 2 h after 2 hours of water bath, the water temperature was adjusted to 75 ° C, and a biotin-DUT, biotin-DATP and ground Gao Xin-dUTP were added to 1u's Kjeno enzyme 15min to terminate.
[0055] S5, then add 1 g of Tris balanced phenol and chloroform-isoprene 24: 1, severe oscillate for 10 s, 10000 r / min for 10 min, and the upper aqueous phase was extracted from the upper aqueous phase.
[0056] S6, adding an equal volume chloroform to the new test tube, mixing 1000r / min for 10 min after mixing, and then transfer the upper water phase to another test tube.
[0057] S7, 1 / 4 volume of NaAc, and 2x volume anhydrous ethanol was added in yet the test tube, and after -20 ° C for 12 h, 12000 r / min was centrifuged for 15 min and the upper cleaner was absorbed, and then 75% at 1 ° C. Ethanol.
[0058] Af...
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