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Transgenic Rodent







  • We can perform entire transgenic project or you can outsource specific phases of the project to us
  • Our unique technology ensures a fast turn around time
  • Main strain available for transgenic production are; C57BL/6 (B6) or FVB for mouse and Sprague-Dawley (SD) and Wistar for Rat, although we can use any per your request
  • Desired final deliverables: Transgenic vector, transgenic embryos, transgenic founders, and others found below
Diagram Transgenic Animal

THE SERVICE PROVIDED BY AMSBIO
SERVICES
DESCRIPTION
Transgenic Strategy Design
  • Design a transgenic strategy that meets your research needs
  • A PCR genotyping strategy will also be designed for founder screening
Transgenic Vector Construction
  • Construct conventional plasmid-based transgenic vectors or, in case of BAC-based transgenes, modify BACs by recombineering as needed (for Rats just plasmid-based)
  • Confirm plasmid vector or BAC modification by restriction digest and sequencing as needed.
Pronuclear Injection
  • Inject transgenic vector or BAC into the pronucleus of fertilized eggs
  • Implant injected eggs into surrogate mothers to obtain offspring
Transgenic Mouse Screening
  • Genotype pups by PCR to identify those carrying the transgene
  • If the project goal is to examine lacZ expression in transgenic embryos, we will genotype embryos using yolk sac and perform lacZ staining on the embryo proper
Prepare DNA for injection and develop genotyping strategy
  • Plasmid DNA is typically linearized for injection
  • BAC DNA is typically injected in unlinearized form
  • PCR-based genotyping strategy will be developed and tested
  • If you provide your own transgenic vector, 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
Generation of Transgenic Founder Mice/Rats
  • We guarantee a minimum of 3 transgenic founders in case of plasmid injection or 2 transgenic founders in case of BAC injection
  • Pups will be genotyped by PCR to identify founder mice
Generation of Transgenic Embryos and Staining of Embryos for lacZ Expression
  • Embryos will be retrieved from surrogate mothers at the stage customers specify (typically within the E10.5-E16.5 window)
  • We guarantee a minimum of 3 transgenic embryos in case of plasmid injection or 2 transgenic embryos in case of BAC injection
  • Embryos will be genotyped by PCR. All embryos, regardless of genotype, will also be fixed and stained for lacZ expression
Breeding Transgenic Line to Homozygosity (Optional for Rat)
  • We will obtain homozygous rats of a transgenic line by first intercrossing heterozygous males and females to obtain offspring
  • We will identify homozygous males & females by progeny test on the offspring
  • They will be crossed to obtained additional homozygous rats
  • We guarantee a minimum of 2 males and 2 females that are homozygous for the transgene
Breeding Founders to Obtain F1 (Optional for Mouse)
  • We can breed the founders the obtain F1 mice
  • Breed up to 5 founders to wild type mice of matching strain background, and genotype their offspring to obtain F1 mice bearing the transgene


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