eagle-i The University of PennsylvaniaThe University of Pennsylvania
See it in Search
This page is a preview of the following resource. Continue onto eagle-i search using the button on the right to see the full record.

Mouse DNA microinjection

eagle-i ID

http://eagle-i.itmat.upenn.edu/i/0000016d-d5db-1675-eaa8-15d780000000

Resource Type

  1. Material modification service

Properties

  1. Fee for service
    Yes
  2. Resource Description
    Your DNA will be injected directly into the pronucleus of a minimum of 150 fertilized 0.5d.p.c. zygotes. This can be performed in a wild-type strain, or existing mutant strain. The DNA is loaded into the ultra-fine injection needle, and using a constant, regulated flow of pressure is injected into each zygote. Following an incubation period, the surviving zygotes are surgically transferred into the reproductive tract of 0.5day pseudo-pregnant surrogate females. When the offspring reach 10-14 days old they will be tail snipped, the tissues will be delivered to your lab and screened for integration of the exogenous DNA sequence. Mice identified with the correct sequence are considered the founders, or F0, these will be transferred to your animal holding room, where they can be bred to expand the line for experimental use. The facility will inject your DNA construct on a first-come, first-served basis. We will attempt to accommodate any strain background requests, but please be aware that hybrid strains tend to work better than inbred strains. A minimum of 100 microinjected zygotes are transferred into recipient mother mice, the majority of which will become pregnant. We expect to generate 30-50 pups from a microinjection session, this number is typically sufficient for the production of founder mice. Although it is common that founder mice can be generated from fewer animals too. If the core does not generate any founders from the first microinjection session a second session will be scheduled, the cost of the additional animals will be billed to you, but the core will absorb the service fee. If, however, no founders are identified from the second set of microinjections we will need to arrange a meeting with the principle investigator to discuss the project. PREPARATION OF TRANSGENE DNA FOR MICROINJECTION General Considerations DNA samples for microinjection should be free of contaminants that might harm the single cell zygotes. Such potential contaminants include traces of phenol, ethanol or enzymes. It is also essential to remove any particles that could clog the injection needles. When purifying the DNA, please use powder-free gloves to avoid particulate matter that may interfere with the microinjection. Be aware that autoclaved tubes can have residue in them- ; use sterile cryovials as an alternative. The Transgenic Core Facility recommends that all solutions used to prepare DNA, be filtered through a 0.2micron filter. Sterile, embryo-tested water (Sigma W-1503, 500ml) or Milli-Q should be used in all solutions. Once you have isolated your injection construct and diluted it to the correct concentration for injection, a final pass through a 0.2micron syringe filter (Millipore, Cat. No. SLGVL04) will remove any remaining impurities that may clog the injection needle. This step is essential.
  3. Service Fee URL
    https://corelabs.research.chop.edu/sites/default/files/inline-files/Transgenic_Fee_Schedule_9.25.19.pdf
  4. Service Provided by
    Transgenic Core (CHOP)
  5. Website(s)
    https://corelabs.research.chop.edu/transgenic/services
 
RDFRDF
 
Provenance Metadata About This Resource Record
  1. workflow state
    Published
  2. contributor
    ggrant (Gregory Grant)
  3. created
    2019-10-16T14:38:26.328-04:00
  4. creator
    ggrant (Gregory Grant)
  5. modified
    2019-10-16T14:38:40.762-04:00
Copyright © 2016 by the President and Fellows of Harvard College
The eagle-i Consortium is supported by NIH Grant #5U24RR029825-02 / Copyright 2016