Variable-Cycle PCR Amplification for Uniform DNA Concentrations
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Solution Overview
Problem
Existing PCR methods for nucleic acid tag-based display technologies face inefficiencies due to variability in nucleic acid concentrations after the PCR step, necessitating separate PCR devices or inefficient grouping, which is costly and space-intensive.
Innovation Solution
A method and system for DNA amplification using PCR that includes a cycle number acquisition step, a minimum PCR step, a first transfer step, and a repeated additional PCR/transfer step, utilizing a PCR device with a multiwell plate, thermal cycler, and processor to maintain DNA concentrations within a specific range.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If multiple PCR devices are used to process different DNA samples simultaneously, then processing efficiency and productivity are improved, but device complexity, cost, and space requirements increase
Solution Approach 1:
The invention segments the PCR process into two distinct phases: a common initial PCR step that all samples share, and individual subsequent PCR steps that each sample performs separately. This segmentation allows multiple samples to be processed efficiently using a single PCR device, resolving the contradiction between productivity and device complexity
Solution Approach 2:
The invention performs preliminary PCR amplification for all DNA samples simultaneously in a common initial PCR step before individual samples require separate processing. This preliminary action optimizes resource utilization and enables efficient batch processing, improving productivity without requiring multiple PCR devices
2Stability of the object's composition
If PCR cycle numbers are optimized for each individual sample, then DNA concentration uniformity is improved, but processing time and operational complexity increase
Solution Approach 1:
The invention determines and performs the common initial PCR step for all samples before individual samples require different cycle numbers. This preliminary determination of baseline cycle numbers allows subsequent individualized processing to be more efficient, maintaining concentration uniformity while reducing total processing time
Solution Approach 2:
The invention implements a dynamic PCR processing scheme where samples transition from a common initial PCR phase to individualized subsequent PCR phases with customized cycle numbers. This dynamic approach allows optimization of DNA concentration uniformity while minimizing processing time through flexible, sample-specific cycle adjustment
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
Ensures uniform DNA concentrations across multiple samples, enabling efficient processing without contamination, reducing the need for multiple PCR devices, and preserving DNA diversity for subsequent stages.
Implementation Method 1
a minimum PCR step in which PCR is performed for the minimum cycle number; an additional PCR step in which PCR is performed up to the maximum cycle number
Data Source
Figure 1A
Figure 1B
Figure 1C~2
AI summary
[Problem] To provide a method and a system for amplifying DNA using PCR that can maintain the DNA concentration of a plurality of DNA samples within a certain range. [Solution] The problem is solved by a DNA amplification method using PCR, the method comprising: a cycle number acquisition step in which the number of PCR cycles for each of DNA samples housed in individual wells of a first multiwell plate is acquired; a minimum PCR step in which PCR for the minimum number of cycles, which is the smallest number of cycles obtained in the cycle number acquisition step, is performed; a first transfer step in which a sample that has been identified with the minimum number of cycles, which is the smallest number of cycles found in the cycle number acquisition step, and that has been subjected to PCR for the minimum cycles is transferred to a second multiwell plate; and a step for repeating additional PCR/transfer steps, in which, after the first transfer step, PCR is performed up to a maximum number of cycles, which is the greatest number of cycles acquired in the cycle number acquisition step, and the sample that has reached the number of cycles acquired in the cycle number acquisition step is transferred to the second multiwell plate.