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Nuclease detection method based on protamine assistance and kit

A protamine and detection method technology, applied in the field of nuclease detection, can solve the problems such as difficulty in realizing high-throughput and automation of nuclease, introduction of radioactive substances, complex modification process, etc. Effect

Inactive Publication Date: 2020-02-11
YANTAI UNIV
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AI Technical Summary

Benefits of technology

This patented technology allows for better ways to detect small amounts of DNA that may indicate disease states earlier than other methods such as histological analysis or immunofluorescence techniques. It uses specialized materials called prion proteins made up from different types of protein molecules like enzymes involved in cellular metabolism during growth process. These Prions have specific properties allowing them to bind specifically to certain parts of their target gene sequence. By measuring this binding activity, researchers aimed at understanding how these targets function over time could help identify conditions associated with inherited disorders more easily and accurately.

Problems solved by technology

This patented technical problem addressed in this patents relates to accurately analyzing nucleic acid breakdown caused by certain types of proteins like histones, which makes it hard to analyze them with current techniques due to their complexity and difficulty handling.

Method used

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  • Nuclease detection method based on protamine assistance and kit
  • Nuclease detection method based on protamine assistance and kit
  • Nuclease detection method based on protamine assistance and kit

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Embodiment 1

[0031] Detection of S1 nuclease (S1 nuclease). S1 nuclease is an endonuclease that can degrade single-stranded DNA and single-stranded RNA to produce 5'-single-stranded nucleotides or oligonucleotides.

[0032] The steps for detecting S1 nuclease using the method provided by the present invention are as follows:

[0033] Step1: Synthesize negatively charged gold nanoparticles and dilute to an absorbance value between 0.2-0.8.

[0034] The Frens method (one-step synthesis method) is used to synthesize negatively charged gold nanoparticles with chloroauric acid as the original material and sodium citrate as the reducing agent. Specifically:

[0035] Boil 100mL of 1mM chloroauric acid solution, and then quickly add 10mL of 38.8mM sodium citrate solution to the boiled chloroauric acid solution. At this time, the color of the solution gradually changes from yellow to light gray, and finally changes to It is wine-red, continue to boil for 30 minutes, stop heating, stir and cool to room tem...

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Abstract

The invention discloses a nuclease detection method based on protamine assistance and a kit. The nuclease detection method comprises the steps of: synthesizing gold nanoparticles with negative charges; optimizing the concentration of a protamine solution enabling the gold nanoparticles to be just agglomerated and the concentration of an aptamer solution enabling the gold nanoparticles to be just not agglomerated; sequentially reacting a nuclease standard solution with the optimized aptamer solution, the optimized protamine solution and a gold nanoparticle solution, and drawing a standard working curve of nuclease; adding to-be-detected samples into the system under the same condition to generate sample signals; and comparing the sample signals with the standard working curve of nuclease toobtain the concentration of nuclease in the to-be-detected samples. The nuclease detection method and the kit have the advantages of simplicity, rapidness, sensitivity, wide detection range and low detection cost, can realize high-throughput and automatic detection of nuclease, and have extremely high application value for early detection of various early diseases.

Description

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Claims

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Application Information

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Owner YANTAI UNIV
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