Genotyping Methods
PCR Based Genotyping
PCR genotyping provides rapid, cost effective verification of targeted alleles.
- Allele Specific PCR: Detects presence of targeted allele
- Wild Type PCR: Confirms absence of wild type allele
- Heterozygous/Homozygous Discrimination: Distinguishes genotype
- Multiplex PCR: Simultaneous detection of multiple alleles
PCR genotyping is ideal for routine colony maintenance and rapid screening of large numbers of animals.
Southern Blot Analysis
Southern blot provides definitive verification of complex allele structures.
- Correct Integration: Confirms targeting at intended locus
- Single Copy Verification: Ensures no random insertions
- Allele Structure: Verifies complete allele architecture
- Pre Germline Characterization: ES cell clone verification
Quantitative PCR (qPCR)
qPCR enables copy number determination and expression analysis.
- Copy Number Analysis: Determines transgene copy number
- Expression Level Quantification: Measures gene expression
- Relative Quantification: Compares expression between samples
- High Throughput Screening: Rapid analysis of multiple samples
Sequencing Based Genotyping
Sequencing provides nucleotide level verification.
- Point Mutation Verification: Confirms specific nucleotide changes
- Small Insertion/Deletion Detection: Identifies small indels
- Fusion Junction Verification: Confirms correct fusion sequences
- Complete Allele Sequencing: Full sequence verification
Sequencing is essential for verifying point mutations and small genetic modifications.
Genotyping Applications
Colony Maintenance
Routine genotyping ensures correct colony composition:
- Breeding Pair Selection:Verify genotypes before mating
- Offspring Screening:Identify desired genotypes
- Colony Purity:Maintain genetic background integrity
- Allele Transmission:Track allele inheritance
Experimental Design
Genotyping supports experimental planning:
- Genotype Verification:Confirm experimental groups
- Control Selection:Identify appropriate controls
- Cohort Assembly:Assemble study cohorts with correct genotypes
- Phenotype Correlation:Link phenotypes to genotypes
Quality Assurance
Genotyping provides quality control:
- Allele Integrity:Verify allele structure over generations
- Contamination Detection:Identify genetic contamination
- Background Verification:Confirm genetic background
- Archival Verification:Verify archived samples
Service Workflow
Sample Submission
Samples can be submitted in multiple formats:
- Tail Biopsies: Standard tail snip samples
- Ear Punches: Alternative tissue samples
- Blood Samples: For specific applications
- ES Cell Samples: For pre germline characterization
Analysis and Reporting
Genotyping results are provided with:
- Clear Genotype Calls: Heterozygous, homozygous, or wild type
- Quality Metrics: Confidence scores for results
- Gel Images: Visual confirmation when applicable
- Interpretation Guidance: Help with result interpretation
Custom Genotyping Development
Assay Design
ingenious targeting laboratory can design custom genotyping assays for:
- Novel Alleles:New targeting projects
- Complex Alleles:Multi component alleles
- Optimized Protocols:Improved efficiency or specificity
- Multiplex Assays:Simultaneous detection of multiple alleles
Protocol Optimization
We optimize genotyping protocols for:
- High Throughput:Rapid screening of large colonies
- Cost Effectiveness:Reduced reagent costs
- Reliability:Improved accuracy and reproducibility
- Specificity:Reduced false positive rates
What Researchers Say
“We found iTL's service to be exceptionally efficient and very well managed. We generated several mouse lines with iTL, and every step for each project was performed quickly and precisely, without any delays or problems.”
— Santos Franco, PhD
University of Colorado Anschutz Medical Campus
Request Genotyping Services
Our team can help you determine the optimal genotyping approach for your mouse model. Contact us to discuss your genotyping requirements.
Frequently Asked Questions
Genotyping Service Insights
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