eagle-i Oregon Health & Science UniversityOregon Health & Science University
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Generating MEFs protocol

eagle-i ID

http://ohsu.eagle-i.net/i/0000012f-7a3b-be79-f561-6e8a80000000

Resource Type

  1. Protocol

Properties

  1. Resource Description
    "Embryos are harvested from naturally mated or superovulated and mated females between e12.5 and e15.5 with the optimal time being e13.5. Heads and internal organs of the embryos are removed and the remaining tissue is rinsed in sterile PBS and minced. After the addition of trypsin and DNAseI the tissue is incubated for 15 minutes and then passed through a 16G needle. The resulting mixture is centrifuged at roughly 1000RPM for 5 minutes and the supernatant is collected and spun again and the very loose pellet of cells is plated in MEF media. The supernatant is allowed to incubate for another 10-15 minutes, spun down and the resulting loose pellet is plated."
  2. Additional Name
    Generating mouse embryo fibroblasts protocol
  3. Uses
    Cook Medical benchtop embryo incubator
  4. Uses
    VWR CO2 embryo incubator
  5. Uses
    Genevac Bio-Cool programmable cryopreservation freezer
  6. Uses
    BTX ECM 830 electroporator
  7. Uses
    Electroporation
  8. Uses
    Establishing cell culture
  9. Used by
    Transgenic Mouse Model Shared Resource
  10. Website(s)
    http://www.ohsu.edu/xd/research/research-cores/transgenics/protocols.cfm
 
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The eagle-i Consortium is supported by NIH Grant #5U24RR029825-02 / Copyright 2016