Each full transgenic project is split into the following stages:
1. Transgenic strategy design
2. Transgenic vector construction
3. Pronuclear injection
4. Transgenic rat screening
Prepare DNA for injection and develop genotyping strategy
The success of transgenic production depends critically on the quality of DNA used in pronuclear injection. Accordingly, we employ a special protocol to prepare top-quality DNA for our injections. Plasmid DNA is typically linearized for injection. BAC DNA is typically injected in unlinearized form as this has been shown to produce similar biological outcome as linearized BAC. PCR-based genotyping strategy will be developed and tested. If you provide your own transgenic vector rather than having us construct it, you can send us either E. coli or unlinearized DNA solution (≥50 µg total), and we will perform the necessary preparation to purify/linearize the DNA. We strongly discourage customers from sending us linearized, injection-ready DNA, because the quality of such DNA is often insufficient for pronuclear injection. If you provide your own vector without sequence information, you will also need to provide a PCR-based genotyping strategy along with your vector.
Generation of transgenic founder rats
- Plasmid injection to obtain founders, basic service: A minimum of 200 eggs will be injected to produce live animals. This will result in an average of 2-3 transgenic founders, but the actual numbers could fluctuate significantly and it is possible that no founders are obtained. If you wish to ensure the generation of founders, we strongly recommend that you use guaranteed service (see below).
- Plasmid injection to obtain founders, guaranteed service: We guarantee a minimum of 3 transgenic founders (the actual number of founders obtained is typically higher). We will inject as many eggs as needed to fulfill this guarantee.
- BAC injection to obtain founders, basic service: A minimum of 200 eggs will be injected to produce live animals. This will result in an average of 2 transgenic founders, but the actual numbers could fluctuate significantly and it is possible that no founders are obtained. If you wish to ensure the generation of founders, we strongly recommend that you use guaranteed service (see below).
- BAC injection to obtain founders, guaranteed service: We guarantee a minimum of 2 transgenic founders (the actual number of founders obtained is typically higher). We will inject as many eggs as needed to fulfill this guarantee.
- Genotyping pups to identify founders: Pups will be genotyped by PCR to identify founder rats.
Breeding transgenic line to F1
For most projects, the generation of founder transgenic rats is the end point. However, you may choose to have us breed the founders further to obtain F1 rats. We will breed up to 3 founders to wildtype rats of matching strain background and genotype their offspring to obtain F1 rats bearing the transgene.
Sprague-Dawley (SD) and Long Evans strains are available for transgenic rat production. Please inquire availability of other strains.
All transgenic rat generations consist of the following production stages. You can contract Cyagen for full service or may also outsource your project to us at any stage.
|1||Transgenic strategy design||Free||1-4 days|
|2||Transgenic vector construction or BAC modification*||Please inquire||2-5 weeks|
|3||Prepare DNA for injection and develop genotyping strategy||$350||1-2 weeks|
|4||Plasmid injection to obtain founders, Basic Service||SD||$9,950||7-11 weeks|
|Long Evans||$10,950||11-15 weeks|
|Plasmid injection to obtain founders, Guaranteed Service||SD||$14,950||7-15 weeks|
|Long Evans||$15,950||11-19 weeks|
|BAC injection to obtain founders, Basic service*||SD||$10,950||10-18 weeks|
|Long Evans||$11,950||17-25 weeks|
|BAC injection to obtain founders, Guaranteed Service*||SD||$15,950||10-25 weeks|
|Long Evans||$16,950||17-29 weeks|
|5||Genotyping pups to identify founders||$250||1 week|
|6||(Optional) Breeding founders to obtain F1||$3,150||9-11 weeks|
* If you need your BAC modified, click here to find out about BAC modification services.
Note: For transgenic rat services not listed above, please inquire about availability and pricing. The turnaround time above does not include the time for obtaining host institution’s approval for rat importation and transit time during shipping.
Cyagen offers the best guarantee in the industry – we will fully refund the client’s service fee if animals with the specified genotype are not generated (except for genetic modifications severely affecting viability, mobidity, or fertility). Given the complexity of biological systems, a particular genetic modification may not result in the desired phenotype. As such, Cyagen's guarantee covers the creation of animals with the specified genotype, not a particular phenotypic outcome in terms of transcription, protein/RNA function, or organismal biology.
If you find another commercial service provider that offers better pricing than ours, we will match the price plus an additional 5% off.
Standard payment terms include a 50% upfront payment before the project, and the remaining 50% plus shipping charge paid after completion of the project. If you need us to design your transgenic strategy, we will provide this service for free irrespective of whether you end up choosing us for your project.
We offer up to 10% bulk discount for large orders. Large orders are defined as 5 or more projects from the same institution. If you bundle your order with that of your colleagues, you can all qualify for the bulk discount.
Products are shipped from our Santa Clara, California facility. For rat shipments, the shipping charge includes courier cost plus a $100/crate handling fee. DNA constructs in E. coli are shipped at room temperature, and the charge includes courier cost plus a $10 handling fee. We typically use World Courier to ship live rats and FedEx for other shipments.
All animal work is conducted in our specific pathogen free (SPF) facilities that have been AAALAC accredited and OLAW assured. For details information, please visit our support section for Description of our Facility, Animal Health and Animal Welfare Program.
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