Method for detecting miRNA-21 through carboxylated graphene oxide modified electrodes

A miRNA-21, modified electrode technology, applied in the detection field, can solve the problems of difficult to develop miRNA, high sequence similarity, low abundance, etc., to achieve the effect of enhancing current intensity and surface area, stabilizing electrochemical signals, and improving accuracy

Active Publication Date: 2019-06-07
宁波远志立方能源科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the short length, high sequence similarity and low abundance (as low as a few molecules per cell) of miRNAs make it difficult to develop fast, simple, sensitive and selective electrochemical methods for the detection of miRNAs

Method used

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  • Method for detecting miRNA-21 through carboxylated graphene oxide modified electrodes
  • Method for detecting miRNA-21 through carboxylated graphene oxide modified electrodes
  • Method for detecting miRNA-21 through carboxylated graphene oxide modified electrodes

Examples

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Embodiment 1

[0033] A method for detecting miRNA-21 at a carboxylated graphene oxide modified electrode, comprising:

[0034] Step 1. Electrode modification: a. Drop the miRNA-21 aptamer solution with a concentration of 2 μM onto the surface of the gold electrode, place it in an environment at 4°C for 12 hours, wash it with a PBS solution with a pH of 7.4 for 25 seconds, and wash it with ultrapure water Rinse well and blow dry with nitrogen;

[0035] b. Add the mercaptoethanol solution with a concentration of 1mM dropwise to the surface of the gold electrode in step a until the surface of the gold electrode is covered with the mercaptoethanol solution. After placing it at 4°C for 1 hour, wash it with a PBS solution with a pH of 7.4 for 25 seconds. Rinse with ultrapure water and blow dry with nitrogen;

[0036] c. Add the reaction solution dropwise to the surface of the gold electrode in step b, place it in an environment of 55°C for 125 minutes, rinse it with a PBS solution with a pH of 7...

Embodiment 2

[0051] The method for detecting miRNA-21 at a carboxylated graphene oxide modified electrode is the same as in Example 1, wherein, it also includes: before step 1, the gold electrode is subjected to the following treatment:

[0052] Place the gold electrode in NaHCO 3 In an ultrapure aqueous solution with a concentration of 5 μM and a concentration of KCl of 10 μM, soak for 40 minutes in a water bath at a temperature of 65 ° C, rinse with ultrapure water, and then place it in an ultrapure aqueous solution with a concentration of acetic acid of 5 μM and a concentration of silicic acid of 2 μM , soaked in a water bath at a temperature of 80°C for 60 minutes, rinsed with ultrapure water, polished with 0.05 μm polishing powder on a polishing cloth, and rinsed with ultrapure water.

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Abstract

The invention discloses a method for detecting miRNA-21 through carboxylated graphene oxide modified electrodes. The method includes the steps that 1, an miRNA-21 aptamer solution is dropwise added tothe surface of a gold electrode, then the gold electrode is flushed and blow-dried, a mercaptoethanol solution is dropwise added, then the gold electrode is flushed and blow-dried, a reaction solution is dropwise added, then the gold electrode is flushed and blow-dried, the gold electrode is immersed in a carboxylated graphene oxide compound saturated solution and then flushed and blow-dried, methylene blue is dropwise added, then the gold electrode is flushed and blow-dried to obtain a modified electrode, ferrocene and diphenylalanine are dissolved in ethyl alcohol, carboxylated graphene oxide is added for ultrasonic treatment, and filtrate is dried to obtain a carboxylated graphene oxide compound; 2, a plurality of modified electrodes are prepared, an impedance value corresponding to each concentration is detected, and the linear relation between difference values of the impedance values and the concentrations is obtained through calculation; and 3, the modified electrode for detecting a to-be-detected sample liquid is prepared, the impedance value of the to-be-detected sample liquid is detected, and the concentration of miRNA-21 in the to-be-detected sample liquid is obtained through calculation according to the linear relation. The method has the beneficial effect of electrochemical signals of the gold electrode are enhanced.

Description

technical field [0001] The present invention relates to the field of detection. More specifically, the present invention relates to a method for detecting miRNA-21 at a carboxylated graphene oxide modified electrode. Background technique [0002] MicroRNAs (miRNAs) are a class of small non-coding RNA molecules containing approximately 22 nucleotides that function in the transcriptional and post-transcriptional regulation of gene expression. According to some recent reports, certain types of miRNAs are closely related to human cancers, and miRNA-21 is one of them. Among these different miRNAs, miRNA-21 is considered as a potential biomarker because it is overexpressed in many cancer tissues, such as lung adenocarcinoma, glioblastoma, breast cancer, cervical cancer. Furthermore, by delivering inhibitors to regulate the expression of specific microRNAs, they could also serve as novel therapeutic targets for many diseases. Therefore, there is an urgent need for sensitive micr...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N27/327G01N27/30G01N27/02G01N27/416B82Y30/00B82Y5/00B82Y15/00
Inventor 肖琦黄珊李家文金晓宇
Owner 宁波远志立方能源科技有限公司
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