Method for evaluating interaction effect of two single-stranded nucleic acid molecules
A technology for single-stranded nucleic acid molecules and capture probes, which is applied in the field of evaluating the interaction of two single-stranded nucleic acid molecules, can solve the problems of limited application, detachment, and difficult regeneration of detection, and achieves simple preparation methods, stable performance, and repeatability Good results
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Embodiment 1
[0045] Example 1. Evaluation of the interaction of two single-stranded DNA molecules (complementary base pairing)
[0046] The inventors of the present invention have provided a method for evaluating the interaction of two single-stranded DNA molecules through a large number of experiments. The specific schematic diagram is shown in figure 1: modify a single-stranded DNA molecule with an amino group as a capture probe (such as capture probe 1 in the following examples), fix it on the surface of an aldehyde-modified fiber optic sensor by Schiff base reaction, and use fluorescent labeling for another single-stranded DNA molecule As a signal probe (as hybridization probe 2, hybridization probe 3, random probe 4, aggregation probe 5 in the following examples), the fluorescent signal is detected after the signal probe reacts with the capture probe: if the fluorescent signal is strong, Then two single-stranded DNA molecules interact (such as figure 1 The bases shown in A are comple...
Embodiment 2
[0076] Embodiment 2, evaluate the interaction of two single-stranded DNA molecules (aggregation reaction)
[0077] 1. Preparation of Capture Probe 1 Modified Optical Fiber
[0078] Same as Step 1 of Example 1.
[0079] 2. Aggregation reaction on the surface of optical fiber
[0080] 1. Dissolving the aggregation probe 5 with pH 7.4 and 10 mM PBS buffer solution to obtain a hybridization solution; the concentration of the aggregation probe 5 in the hybridization solution is 20 nM.
[0081] 2. Install the modified optical fiber of capture probe 1 prepared in step 1 into the all-fiber evanescent wave biosensor.
[0082] 3. Take the hybridization solution and pass it into the all-fiber evanescent wave biosensor that has completed step (1) to detect fluorescent signals at different times.
[0083] 4. Regeneration
[0084] After completing step 3, the modified optical fiber of capture probe 1 can be regenerated. The regeneration method is: take the all-fiber evanescent wave bios...
Embodiment 3
[0088] Embodiment 3, reproducibility detection
[0089] 1. Preparation of Capture Probe 1 Modified Optical Fiber
[0090] Same as Step 1 of Example 1.
[0091] 2. Optical fiber surface regenerative reaction
[0092] 1. Dissolving the aggregation probe 5 with pH 7.4 and 10 mM PBS buffer solution to obtain a hybridization solution; the concentration of the aggregation probe 5 in the hybridization solution is 20 nM.
[0093] 2. Install the modified optical fiber of capture probe 1 prepared in step 1 into the all-fiber evanescent wave biosensor.
[0094] 3. Take the hybridization solution and pass it into the all-fiber evanescent wave biosensor that completed step 2 to detect the fluorescent signals at different times.
[0095] 4. Regenerate the all-fiber evanescent wave biosensor after completing step 3.
[0096] The steps of regeneration are as follows: take the all-fiber evanescent wave biosensor after completing step 3, wash it with SDS aqueous solution with pH7.0 and a co...
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