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Photoelectric chemical analysis method for detection of 5-hydroxymethylcytosine based on black titanium dioxide

A photoelectrochemical, black technology, applied in the field of detection of 5-hydroxymethylcytosine, can solve the problems of insufficient attention, complicated operation, separation of the two, etc., to improve the detection sensitivity, the detection method is simple, and the operation is easy. Effect

Inactive Publication Date: 2019-05-03
SHANDONG AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

5hmC was discovered in phage as early as 1952, but it did not attract enough attention at that time
However, due to the structural similarity of 5hmC and 5mC, it is difficult to separate the two with existing bioassays
[0003] At present, the commonly used methods for detecting 5hmC include single-molecule real-time sequencing, thin-layer chromatography, high performance liquid chromatography-mass spectrometry, capillary electrophoresis-mass spectrometry, etc., but these methods generally have expensive instruments, complicated operations, and require professionals. Operation and other disadvantages
However, there is no report on the detection of 5hmC by photoelectrochemical analysis based on black titanium dioxide.

Method used

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  • Photoelectric chemical analysis method for detection of 5-hydroxymethylcytosine based on black titanium dioxide
  • Photoelectric chemical analysis method for detection of 5-hydroxymethylcytosine based on black titanium dioxide
  • Photoelectric chemical analysis method for detection of 5-hydroxymethylcytosine based on black titanium dioxide

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preparation example Construction

[0080] (2) Black TiO 2 Preparation: Weigh 0.5-2g white TiO 2 , 0.5-2g of sodium borohydride was ground and mixed in a mortar. The mixture is put into a porcelain boat, and calcined at 200-400° C. for 0.5-2 hours under a nitrogen atmosphere. The obtained black solid was washed three times with deionized water and absolute ethanol respectively, and dried under vacuum at 60° C. for later use.

[0081] (3) Preparation of amino-modified zinc oxide: Weigh 0.1-1 g of zinc acetate and add it to 50 mL of absolute ethanol, and stir evenly. Then, 10-30 mL of 0.5M NaOH solution was added with stirring. Then the above solution was transferred to a reaction kettle for 4-8 hours at 120-180°C. The product was washed three times with deionized water and absolute ethanol, and then 0.5-1.5 g of the above product was weighed and added to 100 mL of 0.5 M NaOH solution, stirred and refluxed at 60-90° C. for 15-25 h. The product was washed three times with water and added to a mixed solution of...

Embodiment 1

[0100] Example 1: Black TiO 2 preparation of

[0101] Weigh 1g white TiO 2 , 1g of sodium borohydride was ground and mixed in a mortar. The mixture was put into a porcelain boat and calcined at 250 °C for 1 h under nitrogen atmosphere. The obtained black solid was washed three times with deionized water and absolute ethanol respectively, and dried under vacuum at 60° C. for later use.

Embodiment 2

[0102] Embodiment 2: Preparation of amino-modified zinc oxide

[0103] Weigh 0.4g of zinc acetate and add it to 50mL of absolute ethanol, and stir evenly. Then, 25 mL of 0.5M NaOH solution was added with stirring. Then the above solution was transferred to a reaction kettle for 6 hours at 150°C. The product was washed three times with deionized water and absolute ethanol, and then 1 g of the above product was weighed and added to 100 mL of 0.5M NaOH solution, stirred at 70°C and refluxed for 20 h. After the product was washed three times with water, it was added to a mixed solution of ethanol and water (1:1). With stirring, 3 mL of APTES was added followed by reflux for 20 h. Finally, the product was washed three times with water and dried under vacuum at 60°C for further use.

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Abstract

The invention discloses a photoelectric chemical analysis method for detection of 5-hydroxymethylcytosine based on black titanium dioxide. The method is characterized in that firstly, a photoelectricchemical biosensor for detecting 5hmC is constructed, including an electrode, black TiO2, probe DNA, the 5hmC, p-aminophenylboronic acid, PAMAM and amino-modified zinc oxide which are sequentially fixed on the surface of the electrode. The method is advantaged in that photoelectric properties of the black TiO2, the specific catalysis of glycosyltransferase for the 5-hydroxymethylcytosine, the amplification effect of a PAMAM signal and signal quenching of ZnO are utilized to achieve sensitivity detection of the 5-hydroxymethylcytosine. The method is advantaged in that operation is simple, miniaturization is facilitated, and detection of the 5-hydroxymethylcytosine can be realized through only modifying the surface of the ITO electrode.

Description

technical field [0001] The invention relates to the technical field of photoelectrochemical analysis, in particular to a method for detecting 5-hydroxymethylcytosine based on photoelectrochemical analysis of black titanium dioxide. Background technique [0002] 5-Hydroxymethylcytosine (5hmC) is an important epigenetic modifier, which is produced by catalyzed oxidation of 5-methylcytosine (5mC) by TET family enzymes, and is called the sixth base of DNA. 5hmC was discovered in phage as early as 1952, but it did not attract enough attention at that time. It was not until 2009 that researchers discovered that 5hmC was abundantly expressed in mouse, human brain and embryonic stem cells attracted people's attention. As an important epigenetic marker, 5hmC plays a role in many life activities, such as regulating gene activity, gene recombination, cell differentiation, etc. Moreover, studies have found that the content of 5hmC in cancer cells is significantly reduced, so the detec...

Claims

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Application Information

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IPC IPC(8): G01N27/26G01N27/30G01N27/327
Inventor 殷焕顺周云雷艾仕云隋程吉陈燕李菲
Owner SHANDONG AGRICULTURAL UNIVERSITY
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