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A DNase I activity colorimetric assay is provide

A detection method and colorimetric technology, applied in the field of analytical chemistry, can solve problems such as the inability to effectively affect peroxidase activity, and achieve rapid quantification and biological activity detection, and the effect of strong peroxidase activity

Inactive Publication Date: 2019-01-15
NANJING UNIV OF TECH
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Problems solved by technology

Inspired by these reports and our continuing research interest, the present inventors found that single-stranded DNA (ssDNA) of different lengths can effectively regulate the peroxidase activity of MIL-53(Fe), in which the long chain with a longer base number ssDNA has a stronger promoting effect, but short-chain ssDNA with fewer bases cannot effectively affect the peroxidase activity of MIL-53(Fe)

Method used

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  • A DNase I activity colorimetric assay is provide
  • A DNase I activity colorimetric assay is provide

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Experimental program
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Embodiment 1

[0036] (1) With low-cost terephthalic acid and FeCl 3 ·6H 2 O is used as the reaction raw material to synthesize the metal-organic framework material MIL-53(Fe): first, 0.334 g of terephthalic acid and 0.545 g of FeCl 3 ·6H 2 O was dissolved in 10 mL of DMF, and the mixture was stirred at room temperature for 10 min. The resulting solution was transferred to a 50 mL Teflon-lined stainless steel autoclave, which was then sealed and placed in an oven at 150 °C for 17 h.

[0037] (2) After the product in step (1) was cooled to room temperature, the yellow precipitate was collected by centrifugation at 6000 rpm for 3 minutes, and washed thoroughly with deionized water to remove excess reactants. The product was dried under vacuum at 60°C for 24 hours. Finally, the yellow product MIL-53(Fe) was obtained. (3) Prepare the MIL-53(Fe) obtained in step (2) into a 0.1 mg / mL aqueous solution for use, and mix well on a shaker before each use.

[0038] (4) Add 8 μL of 2.5 μM ssDNA (M2...

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Abstract

The invention discloses a colorimetric detection method for deoxyribonuclease I (DNase I) activity based on DNA regulating MIL-53(Fe) peroxidase activity. The invention uses cheap 3, 3 ', 5, 5'-Tetramethylbenzidine (TMB) is a colorimetric substrate, which can regulate MIL-53 (Fe) peroxidase activity and the hydrolysis of ssDNA by DNase I, a biosensor is constructed to observe the activity of DNaseI with naked eyes. A colorimetric method with the advantages of low cost, simple operation and convenience has been developed. This method has high sensitivity and good stability, and is suitable forpopularization and application.

Description

technical field [0001] The invention belongs to the field of analytical chemistry, in particular to a colorimetric detection method for DNase I activity based on DNA regulation of MIL-53(Fe) peroxidase activity. Background technique [0002] DNase I is an endonuclease that can hydrolyze single-stranded or double-stranded DNA to produce single-deoxynucleotides or single-stranded or double-stranded oligodeoxynucleotides. It is widely distributed in mammalian body fluids and tissues, especially the pituitary and pancreas. In addition to its well-known role in pancreatic digestion, DNase I plays an important role in DNA repair, gene recombination and apoptosis. The lack or reduction of DNase I can cause human systemic lupus erythematosus, gastric cancer and colon cancer, while the serum DNase I activity of breast cancer and diabetes patients is higher than normal. In addition, DNase I has been used as a diagnostic marker for cardiovascular diseases such as acute myocardial inf...

Claims

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Application Information

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IPC IPC(8): C12Q1/34
CPCC12Q1/34
Inventor 宋婵丁炜姚玥玮姚成
Owner NANJING UNIV OF TECH
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