Unlock instant, AI-driven research and patent intelligence for your innovation.

A method for evaluating the interaction of two single-stranded nucleic acid molecules

A single-stranded nucleic acid molecule and capture probe technology, which is applied in the field of evaluating the interaction of two single-stranded nucleic acid molecules, and can solve the problems of limited application, detachment, and difficult regeneration of detection.

Active Publication Date: 2021-04-09
TSINGHUA UNIV
View PDF4 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, due to the limitation of the gold electrode surface modification method in the surface plasmon resonance method, it cannot be repeated detection
DNA usually interacts with the gold surface through its surface-modified thiol group, and is adsorbed to the surface of the gold electrode by adsorption method, while the gold thiol group is easily detached from the electrode surface during regeneration, making its detection difficult to regenerate, and a gold electrode can only complete DNA once. Molecular detection limits its application

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • A method for evaluating the interaction of two single-stranded nucleic acid molecules
  • A method for evaluating the interaction of two single-stranded nucleic acid molecules
  • A method for evaluating the interaction of two single-stranded nucleic acid molecules

Examples

Experimental program
Comparison scheme
Effect test

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...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

The present invention discloses a method for evaluating the interaction of two single-stranded nucleic acid molecules. The method is to modify a single-stranded DNA molecule with an amino group as a capture probe and immobilize it on the surface of an aldehyde-modified optical fiber sensor, and use a fluorescent label to another single-stranded DNA molecule as a signal probe, and detect the signal probe after reacting with the capture probe. Fluorescent signal: If the fluorescent signal is strong, the two single-stranded DNA molecules interact; if the fluorescent signal is weak, the two single-stranded DNA molecules do not interact. Therefore, the strength of the fluorescent signal after the reaction between the signal probe and the capture probe can reflect the interaction between the two single-stranded DNA molecules, and the stronger the fluorescent signal, the greater the interaction between the two single-stranded DNA molecules. The preparation method of the invention is simple, the performance is stable, the optical fiber repeatability is good, and the invention has important application value.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a method for evaluating the interaction of two single-stranded nucleic acid molecules. Background technique [0002] At present, the research on the interaction between DNA molecules mainly focuses on the study of the hybridization dynamics between DNA molecules by using the specific complementary pairing rules between DNA molecules. The body structure is less studied. Some DNA intermolecular interaction simulation algorithms, most of them also use some calculation methods related to the minimization of free energy to find the most stable and most likely secondary structure. In addition, there are some methods developed on the basis of minimization of free energy, but all Free energy analysis of base pairing is inseparable. Therefore, having a method capable of analyzing the interactions between DNA molecules, including complementary base pairing, is of great significance in analyz...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6825
CPCC12Q1/6825C12Q2565/607C12Q2563/107
Inventor 何苗唐云飞韩世同
Owner TSINGHUA UNIV