Rapid Nucleic Acid Blotting via Substrate Absorption and Thermal Fixation
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Solution Overview
Problem
Conventional nucleic acid blotting methods are time-consuming and require multiple procedures, making them inefficient for rapid analysis and economically costly, especially for urgent experiments or simple qualitative tests.
Innovation Solution
A rapid blotting method that eliminates the need for prehybridization and reduces the time for nucleic acid probe pairing and washing by using substrates with pores, allowing for quick absorption and fixation of nucleic acid, followed by brief hybridization and detection, significantly shortening the overall process to 15-30 minutes.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Loss of time
If conventional blotting methods are used, then nucleic acid analysis can be performed with standard procedures, but the analysis time is long and material costs are high
Solution Approach 1:
The patent extracts and eliminates the prehybridization step from the conventional blotting procedure. By removing this unnecessary step, the analysis time is significantly reduced while maintaining detection accuracy, directly addressing the contradiction between time loss and productivity
Solution Approach 2:
The patent performs preliminary fixation of nucleic acid to the membrane before hybridization, ensuring that the nucleic acid is properly positioned and secured. This preliminary action eliminates the need for subsequent prehybridization blocking steps, reducing overall analysis time while maintaining detection reliability
2Reliability
If prehybridization is performed to block unfixed areas, then non-specific binding is prevented, but the procedure time and material costs increase
Solution Approach 1:
The patent performs preliminary fixation of nucleic acid to the membrane before hybridization, ensuring that the nucleic acid is properly positioned and secured. This preliminary action eliminates the need for subsequent prehybridization blocking steps, reducing overall analysis time while maintaining detection reliability
Solution Approach 2:
The patent removes the prehybridization blocking step from the conventional protocol. By extracting this unnecessary step, the method maintains detection accuracy through improved fixation techniques while significantly reducing procedure time and material consumption
3Quantity of substance
If membrane is soaked in SSC solution for pretreatment, then nucleic acid absorption is improved, but the time and reagent costs increase
Solution Approach 1:
The patent changes the pretreatment parameters by replacing extended SSC soaking with a brief acid wash step. This parameter change achieves adequate nucleic acid absorption and membrane preparation in much less time, directly reducing pretreatment time while maintaining absorption effectiveness
Solution Approach 2:
The patent uses a simple acid wash solution as a disposable pretreatment step instead of time-consuming SSC soaking. This approach provides sufficient membrane preparation and nucleic acid absorption in a quick, low-cost manner, eliminating the need for prolonged reagent exposure
4Strength
If nucleic acid is fixed by prolonged drying or UV radiation, then fixation strength is improved, but the time and energy consumption increase
Solution Approach 1:
The patent optimizes fixation parameters by using controlled heat treatment at elevated temperatures for a limited duration. This parameter change achieves strong nucleic acid fixation to the membrane in a time-efficient manner, balancing fixation strength with reduced processing time
Solution Approach 2:
The patent utilizes thermal phase transition (heating) to achieve rapid nucleic acid fixation to the membrane. By controlling the temperature and duration of heat treatment, the method achieves strong fixation without requiring prolonged processing or UV radiation, thus reducing time and energy consumption
Applied Scientific Principles
This section explains which scientific principles are used to turn an abstract innovation direction into a practical engineering solution.
Function Achieved in This Case
This method enables rapid nucleic acid analysis by reducing the time and material costs associated with traditional methods, achieving efficient and cost-effective results while minimizing background noise.
Implementation Method 1
transferring a nucleic acid to be analyzed to the substrate and making the nucleic acid to be analyzed absorbed by the substrate
Implementation Method 2
adding a solution containing a nucleic acid probe on the substrate of step (3) to base-pair the nucleic acid probe with the nucleic acid to be analyzed thereon
Data Source
AI summary
The present invention relates to a blotting method for rapidly analyzing nucleic acid comprising the steps of transferring a nucleic acid to be analyzed to the substrate and fixing the nucleic acid to be analyzed absorbed on the substrate; directing adding a nucleic acid probe to hybridize in a short time, without blocking the areas where the nucleic acid to be analyzed has not been fixed; removing the nucleic acid probe which has not been annealed to the nucleic acid to be analyzed by washing; and finally detecting the hybridization signal. According to the present invention, since the prehybridization is not needed and the hybridization and washing time is shortened, the time for the nucleic acid hybridization is dramatically shortened. Therefore, the whole blotting procedures for rapidly analyzing nucleic acid may be finished quickly.