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DNA injection service

eagle-i ID

http://eagle-i.itmat.upenn.edu/i/0000013a-5158-2c61-01af-beb880000000

Resource Type

  1. Material processing service

Properties

  1. Fee for service
    Yes
  2. Resource Description
    "Direct microinjection of DNA into the male pronucleus of a mouse zygote has been the method most extensively used in the production of transgenic mice. If the foreign DNA integrates into the mouse chromosomal DNA at the one-cell stage, the animal will contain the injected DNA in every cell, including those of the germ line. If, however, integration occurs later, the resulting animals will be mosaic for the presence of the injected DNA. The number of copies of integrated DNA can vary from one to several hundred, arranged primarily in tandem head to tail arrays. <strong>What the Facility will Provide</strong> Upon receipt of the DNA construct, proven by documentation to be of high quality and suitable for microinjection (see below), we will proceed to inject a minimum of 150 fertilized eggs. After overnight incubation, normal 2-cell embryos will be transferred into foster females and their pregnancy will be monitored (the gestation period of the mouse is 19-20 days). Two weeks after birth of potential transgenic pups, the project scientist will be notified of the number of pups.The facility will carry out tail clipping, animal tagging and genotyping by PCR according to the conditions set by the project scientist. At weaning, the positive founders will be transferred to the care of the individual investigators as per ULAR procedures.If these studies (with adequate controls) fail to demonstrate the existence of at least one transgenic animal, we will reinject at least 80 eggs at no additional cost to the investigator. If, however, no transgenic animal is detected after the second trial, a meeting will be held between the scientist, the Facility Co-Directors, and the Technical Director to discuss the DNA construct, the tail biopsy analysis, and further plans to be agreed upon. On average, 230 injected eggs should yield at least one (and usually more) transgenic mouse."
  3. Service Fee URL
    http://www.med.upenn.edu/genetics/tcmf/charges.shtml
  4. Additional Name
    DNA microinjection
  5. Additional Name
    Injection of DNA into mouse eggs for the production of transgenic mice or embryos
  6. Contact
    Richa, Jean, Ph.D.
  7. Topic
    Mus musculus
  8. Service Provided by
    Transgenic and Chimeric Mouse Facility (Penn)
  9. Related Publication or Documentation
    DNA purification protocols
  10. Related Publication or Documentation
    Cdx2 levels modulate intestinal epithelium maturity and Paneth cell development
  11. Related Publication or Documentation
    Insulin resistance and a diabetes mellitus-like syndrome in mice lacking the protein kinase Akt2 (PKB beta)
  12. Website(s)
    http://www.med.upenn.edu/genetics/tcmf/services_dna_inject.shtml
  13. Related Technique
    Transgenic mouse model generation
  14. Related Technique
    Microinjection
 
RDFRDF
 
Provenance Metadata About This Resource Record
  1. workflow state
    Published
  2. contributor
    fcoldren
  3. created
    2012-10-11T14:49:23.263-05:00
  4. creator
    fcoldren
  5. modified
    2016-04-21T13:32:29.275-04:00

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The eagle-i Consortium is supported by NIH Grant #5U24RR029825-02 / Copyright 2016