GenoTyping Center of America

Colony Planning and Curation Services

Our colony planning and curation consulting services are backed by extensive colony management, strain development and curation experience. We form a bridge between vivarium staff and investigators, providing strain information, understanding and direction to help prevent costly breeding errors that hinder experimental reproducibility and delay results. Our capabilities include, but are not limited to:

Breeding plan creation
Colony troubleshooting
Managing phenotype expectations
Strain development (from founder to stable transmission)
Curation of newly developed, imported or existing strains (curation is dynamic and can be tailored to your current animal database)
Our Colony Strategy Managers work to understand your specific needs and will then provide a detailed plan with costing for your review. Accept some or all the proposed support services to enhance your existing colony management programs.

Our point mutation genotyping service uses probe based PCR for SNP (Single Nucleotide Polymorphism) and Indel (insertion and deletion) testing for large and small scale projects including SNP, validation, and screening. Our point mutation genotyping service also reports zygosity for targets.

Results are GTCAssured with each sample tested using a single assay with two independent runs. We utilize a proprietary DNA isolation methodology to maximize DNA quality and perform PCR melt curve analysis to generate quality, unambiguous results. Our reports include all methods and raw data for complete evaluation and publication purposes and are delivered in 3-business days guaranteed (allow additional time for new assay development.)

Please provide the following sample submission information:

• Strain name
• Mutation design information (for commercially available strains only the vendor information is necessary)
• Control samples (heterozygous and homozygous, if available)

Our zygosity testing service for transgenic models uses hydrolysis probes to quantitatively determine if samples are homozygous or hemizygous with respect to the transgene of interest. This differs from our standard genotyping service for transgenic samples which only reports qualitative results (presence/absence.) Zygosity is determined by comparison to known controls using the delta Cp (crossing point). Heterozygous and homozygotes are expected to be separated by 1 delta Cp.

Results are GTCAssured with each sample tested using a single assay with two independent runs. We utilize a proprietary DNA isolation methodology to maximize DNA quality and generate quality, unambiguous results. Our reports include all methods and raw data for complete evaluation and publication purposes and are delivered in 3-business days guaranteed (allow additional time for new assay development.)

Please provide the following sample submission information:

• Strain name
• Strain crossing information (to ensure that our assays do not cross react with other mutations)
• Mutation design information (for commercially available strains only the vendor information is necessary)
• Hemizygous and homozygous controls (only needed with first sample submission for assay development)*

*Calls are based on relative amplification vs. the performance of control samples. In the absence of proper controls, tentative calls can be made based on relative amplification if both genotypes are present. However, no zygosity call can be made if only one genotype is present.

We pride ourselves on developing the best assays for your genotyping needs. The more information we have, the better the design. To make a design, we need the following:

• Name of the strain. Your in-house name and the official nomenclature
• Name of the vendor that it came from, if it is commercially available
• Map and/or description of how the mutant was produced
• If it is a transgene, do you plan on matings capable of producing homozygotes?  Do you need to know zygosity?
• Complete sequence, if available
• Current genotyping method and primer sequences
• Known positive controls. Ideally, homozygous, het- or hemizygous, and wild-type.

If you do not have all of the needed information, send us what you have and we may be able to find what we need.  Many commercial vendors provide more than enough information on their websites about their strains.

Our PCR genotyping service offers assays for common transgene targets, such as neomycin, luciferase, GFP, lacZ and Cre recombinase, as well as custom gene-specific assay development for targeted mutations (knock-ins/knock-outs). We design two unique assays for each target at no cost to you. DNA is extracted using our proprietary methods and your samples are tested with the two assays to provide definitive results. For targeted mutations, we determine and report on zygosity for each sample. Transgenic samples have presence/absence reported (designated as wild-type or hemizygous – homozygous are not differentiated.) Zygosity of transgenic samples is determined with our higher level zygosity testing service.

Results are GTCAssured with two independent assays for every target. We utilize a proprietary DNA isolation methodology to maximize DNA quality and perform PCR melt curve analysis to generate quality, unambiguous results. Our reports include all methods and raw data for complete evaluation and publication purposes and are delivered in 2-business days guaranteed (allow additional time for new assay development.)

Please provide the following sample submission information:

• Strain name
• Mutation design information (for commercially available strains only the vendor information is necessary)
• Strain crossing information (to ensure that our assays do not cross react with other mutations)
• Control samples for targeted mutations. No controls necessary for generic assays