Multiplex PCR Genotyping for Mouse Strain Identification
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Solution Overview
Problem
Current methods lack standardized and reliable genetic monitoring for outbred populations of rodents and fish, particularly in identifying and characterizing genetic diversity using autosomal and Y-chromosomal markers, which is essential for maintaining genetic heterogeneity and reproducibility in research.
Innovation Solution
A method involving the simultaneous amplification of multiple polymorphic autosomal and Y-chromosomal STR markers using PCR, allowing for the genetic identification and differentiation of animals, and a kit containing oligonucleotide primer pairs and reagents for multiplex PCR to detect specific alleles, enabling comprehensive biostatistical characterization of outbred populations.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If multiple genetic markers are analyzed using separate PCR reactions, then the accuracy of genetic identification is improved, but the time required and complexity of the procedure increases
Solution Approach 1:
The patent combines multiple separate PCR reactions into a single multiplex PCR reaction that simultaneously amplifies multiple autosomal and Y-chromosomal markers. This merging approach maintains the accuracy of individual marker analysis while reducing the total analysis time and procedural complexity by performing all amplifications in one reaction mixture.
Solution Approach 2:
The multiplex PCR system serves multiple functions simultaneously: it amplifies autosomal markers for individual identification, amplifies Y-chromosomal markers for population differentiation, and provides comprehensive genetic characterization all in one reaction. This multi-functionality resolves the contradiction by achieving high measurement precision across multiple markers without the time penalty of sequential analysis.
2Reliability
If comprehensive genetic monitoring of outbred populations is implemented, then the reliability of experimental data is improved, but the complexity of genetic analysis increases
Solution Approach 1:
The patent segments the complex task of genetic monitoring into distinct marker types (autosomal STR markers for individual identification and Y-chromosomal STR markers for population-level analysis). This segmentation allows comprehensive genetic monitoring to be achieved through a structured approach that manages complexity by organizing analysis into functional categories while maintaining high reliability.
Solution Approach 2:
The patent changes the parameters of genetic analysis by using STR (short tandem repeat) markers with specific repeat units that provide high polymorphism information. This parameter change enables comprehensive genetic monitoring with improved reliability while managing complexity through the use of standardized, highly informative markers that can be analyzed efficiently using the multiplex PCR system.
3Manufacturing precision
If standardized genetic markers are used across different breeding populations, then the comparability of results is improved, but the adaptability to different species and strains decreases
Solution Approach 1:
The patent achieves universality by developing a multiplex PCR system that can simultaneously analyze multiple markers within a species, making the methodology adaptable across different mouse strains and breeding populations. The standardized STR markers and multiplex approach enable comparable results across studies while maintaining versatility through species-specific optimization and the ability to accommodate different strain backgrounds.
Solution Approach 2:
The patent applies local quality by selecting specific STR markers that are polymorphic and informative for particular species or strain groups. This allows standardized analysis within each local context (species or strain) while maintaining overall comparability through the use of consistent methodological approaches. The marker selection is optimized for local genetic characteristics while the multiplex system provides standardized analysis protocols.
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 provides a reliable means to assess and document genetic diversity within outbred populations, ensuring genetic monitoring and quality control, thereby enhancing the reproducibility and reliability of experimental data.
Implementation Method 1
The invention specifically relates to the simultaneous amplification of at least five different polymorphic autosomal markers and at least two polymorphic Y-chromosomal markers of mice in a single reaction using polymerase chain reaction or other multiplex methods
Data Source
Figure 1
AI summary
The invention relates in general to the analysis of genetic markers in individuals of non-human origin, in particular the genotyping of outbred populations of rodents or fish. The invention relates specifically to the simultaneous amplification of at least five different polymorphic autosomal markers and at least one polymorphic Y-chromosomal marker of the mouse in a reaction preparation by means of polymerase chain reaction or other multiplex methods and the detection of the specific alleles for each marker of the multiplex method. The invention further relates to a kit for genetically identifying and/or for differentiating two or more animals from DNA extracts of individuals from wild-type populations, from different inbred or outbred strains, of an identical inbred or outbred strain, or from substrains of an identical inbred or outbred strain, in particular of the rat or mouse.