Nucleic Acid Sample Preparation via Heating and Electrodialysis
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Solution Overview
Problem
Current methods for preparing nucleic acid samples for amplification reactions are complicated, involving multiple steps with nucleic-acid-absorbent porous membranes and require additional enzyme solutions, making the process cumbersome and time-consuming.
Innovation Solution
A method involving a heating step at 50° C. to 65° C. and an electrodialysis step to reduce electrical conductivity to 2,000 μS/cm or less, optionally accompanied by ultrasonic treatment and dilution, simplifies the sample preparation process without the need for pre-prepared enzyme solutions.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Manufacturing precision
If a nucleic-acid-absorbent porous membrane method is used for purification, then nucleic acids can be purified to a suitable level, but the purification operation becomes complicated with multiple steps
Solution Approach 1:
The invention extracts and removes inhibitory substances (proteins, metabolites, salts) from the nucleic acid sample through heating and centrifugation steps, separating the problematic components from the nucleic acids without requiring complex membrane-based purification procedures
Solution Approach 2:
The heating step serves multiple functions simultaneously: it denatures and precipitates proteins, inactivates nucleases, and facilitates the separation of inhibitory substances, combining several purification functions into a single operation that simplifies the overall process
2Reliability
If enzyme solutions are added to reduce inhibition effects, then amplification inhibition is reduced, but the operation becomes complicated requiring pre-prepared enzyme solutions
Solution Approach 1:
The heating step enables the sample to self-purify by denaturing and precipitating inhibitory proteins and metabolites through thermal treatment alone, without requiring external enzyme solutions or additional reagents, making the process simpler and more self-contained
3Manufacturing precision
If multiple purification steps are performed, then nucleic acid purity is improved, but the time required for preparation increases
Solution Approach 1:
The invention combines protein denaturation, nuclease inactivation, and inhibitory substance removal into a single heating step followed by centrifugation, achieving multiple purification objectives simultaneously and reducing the total preparation time compared to sequential purification steps
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 approach simplifies the sample preparation for nucleic acid amplification reactions, reducing the complexity and time required while effectively reducing inhibitory substances, thereby enhancing the efficiency of the amplification process.
Implementation Method 1
a heating step of applying heat to a nucleic acid-containing sample
Implementation Method 2
an electrodialysis step of bringing an electrical conductivity of the sample to 2,000 μS/cm or less
Implementation Method 3
a step of passing a sample solution containing nucleic acids through a nucleic-acid-absorbent porous membrane
Data Source
AI summary
There is provided a method for preparing a sample for a nucleic acid amplification reaction, the method including a heating step of applying heat to a nucleic acid-containing sample, and an electrodialysis step of bringing an electrical conductivity of the sample to 2,000 μS/cm or less.