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Method for detecting microRNA content of to-be-detected liquid

A technology for liquid and content to be tested, applied in the field of electrochemical detection, can solve the problems of high detection cost, small dynamic range, manual operation, etc., and achieve the effects of improving sensitivity and accuracy, reducing background signal, and maintaining specificity

Active Publication Date: 2015-04-29
SUZHOU INST OF BIOMEDICAL ENG & TECH CHINESE ACADEMY OF SCI
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  • Claims
  • Application Information

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Problems solved by technology

[0003] At present, the methods for detecting microRNA mainly include Northern blot analysis, microarray analysis, and real-time quantitative PCR, etc., but these methods all require large, complex, and expensive equipment support, and some even require a large number of manual operations, and the dynamic range is small. The detection cost is high and it is not suitable for routine analysis and detection. Therefore, it is necessary to develop a method that is convenient, simple, low-cost, sensitive and accurate

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  • Method for detecting microRNA content of to-be-detected liquid
  • Method for detecting microRNA content of to-be-detected liquid
  • Method for detecting microRNA content of to-be-detected liquid

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Embodiment 1

[0038] The target microRNA sequence in the test solution is: 5'-UAGCUUAUCAG ACUGAUGUUGA -3'.

[0039] Therefore design:

[0040]The sequence of the first single-stranded DNA molecule is: 5'-P-CTGATAAGCTATTTT-SH-3';

[0041] The sequence of the second single-stranded DNA molecule is: 5'-NH 2 -TTTT TCAACATCAGT -3'.

[0042] Wherein, it should be noted that the underlined part in the target microRNA sequence is complementary to the underlined part in the sequence of the second single-stranded DNA molecule, and the italic part in the target microRNA sequence is complementary to the italic part in the sequence of the first single-stranded DNA molecule . The method for synthesizing two single-stranded DNA molecules is not limited, and is a conventional technique, and an ABI3900 model DNA synthesizer can be used to synthesize any sequence according to the design.

[0043] The synthesized first single-stranded DNA molecule was modified on the surface of the gold electrode by d...

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Abstract

The invention discloses a method for detecting the microRNA content of to-be-detected liquid, which comprises the following steps that: 1, molecules of a first single-stranded DNA are modified on the surface of a working electrode of an electrochemical device; 2, a sample is added, wherein a target microRNA in the sample and the first single-stranded DNA are in partial complementary pairing so as to form a double-stranded structure; 3, a second single-stranded DNA marked with electrical single molecules is added, and due to the existence of partial complementarily-paired sequences between the second single-stranded DNA and the microRNA, a hybrid structure of DNA / microRNA / DNA is formed; 4, the first DNA and the second DNA are connected to be a single-stranded DNA by a DNA ligase, and by heated denaturation, the microRNA is removed from the surface of the electrode so as to effectively reduce background signals; and 5, by detecting electrical signal molecules at the tail end of the DNA, whether the sample contains the target microRNA and corresponding concentrations of the target microRNA can be judged.

Description

technical field [0001] The invention relates to the field of electrochemical detection, in particular to an electrochemical method for detecting the content of microRNA in a liquid to be tested. Background technique [0002] MicroRNA is a kind of single-stranded small molecule RNA expressed endogenously in the body, with a length of about 18-25 nucleotides, located in the non-coding region of the genome, and does not have an open reading frame itself. Mature microRNAs repress the expression of target genes at the post-transcriptional level by complementary binding to mRNAs. microRNA is ubiquitous in eukaryotic cells, and the number is considerable, accounting for about 2% of the total number of genes in the entire genome. microRNA is highly conserved, time-sequential and tissue-specific. microRNA has been found to play an important role in various physiological processes such as timing development, cell apoptosis, fat metabolism, neuron development, cell differentiation, an...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
Inventor 缪鹏王弼陡王磊唐玉国
Owner SUZHOU INST OF BIOMEDICAL ENG & TECH CHINESE ACADEMY OF SCI
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