Electrochemical detection method of micro ribonucleic acid

A detection method and electrochemical technology, applied in biochemical equipment and methods, microbial determination/inspection, etc., can solve the problems of high detection cost, unsuitable for analysis and detection, etc., and achieve strong anti-interference ability, simple operation and application prospect. wide range of effects

Active Publication Date: 2014-08-20
SUZHOU INST OF BIOMEDICAL ENG & TECH CHINESE ACADEMY OF SCI
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] Most of the above methods need the support of large and expensive equipment, and the detection cost is high, so they are not suitable for routine analysis and detection. Therefore, it is necessary to develop a simple, low-cost and sensitive detection method

Method used

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  • Electrochemical detection method of micro ribonucleic acid
  • Electrochemical detection method of micro ribonucleic acid
  • Electrochemical detection method of micro ribonucleic acid

Examples

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Embodiment 1

[0043] 1) Determine the target microRNA sequence according to the measured sample as UAGC UUAU CAGA CUGA UGUU GA; (3'-5')

[0044] 2) Synthesize blocking DNA according to the target microRNA sequence to be detected, the sequence is: AGAA TCTC AACA TCAG TCTG ATAA GCAT; (3′-5′)

[0045] 3) Synthesize reporter DNA according to the blocked DNA sequence: Fc-TAT CAGA CTGA TGTT GAGA TTCT GATA-(C)6-SH-(C) 6 ;(3'-5')

[0046] 4) Primer DNA: GAGA TTCT (3'-5')

[0047] 5) The synthesized reporter DNA molecules were dissolved in a solution containing 10 mM Tris-HCl, 10 mM TCEP, 1 mM EDTA, and 0.1 M NaCl, with a final concentration of 1 μM. Soak the gold electrode in the above solution and let it stand overnight. Then rinse the gold electrode and soak it in 1mM mercaptohexanol for half an hour; then soak the gold electrode in a solution of blocking DNA molecules at a concentration of 1 μM and let it stand for 1 hour to form double-stranded a and open the stem-loop structure of the repor...

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Abstract

The invention discloses an electrochemical detection method of trace micro ribonucleic acid, which comprises the steps that the report DNA is fixed on the electrode surface and forms double strands (a) with block DNA; the block DNA is in molecular combination with the target micro RNA to form double strands (b) so that the block DNA is separated from the electrode surface; the primer DNA is combined with the end sequence of the block DNA in the double strands (b) and self-replicated to form DNA double strands (c), and the target micro RNA is replaced; the replaced target micro RNA returns to the working electrode surface and reacts with other double strands (a); and after N circulating reactions, the report DNAs of the stem-loop structure on the electrode surface are increased, and the electrochemical signal becomes stronger so as to realize electrochemical detection on the monomolecular micro RNA. The same principle also can be promoted to the detection of DNA and siRNA, and the application prospect is wide.

Description

technical field [0001] The invention relates to the technical field of small ribonucleic acid detection, in particular to an electrochemical single-molecule detection method for trace small ribonucleic acid. Background technique [0002] MicroRNA (microRNA) is a kind of small molecule RNA with a length of about 21-25 nucleotides, which inhibits the expression of target genes at the post-transcriptional level by complementary binding with mRNA (messenger RNA). MicroRNA is ubiquitous in multicellular organisms , and the number is considerable, accounting for about 2% of the total number of genes in the entire genome. microRNA is highly conserved in the process of biological evolution, and has been confirmed to participate in and regulate a variety of physiological processes including timing development, cell apoptosis, fat metabolism, neuron development, cell differentiation, hormone secretion, etc., as well as lung cancer, Various diseases including leukemia. Researchers mo...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
CPCC12Q1/6844C12Q2537/1373C12Q2565/607
Inventor 韩坤缪鹏王弼陡唐玉国
Owner SUZHOU INST OF BIOMEDICAL ENG & TECH CHINESE ACADEMY OF SCI
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