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Generating MEFs protocol
eagle-i ID
http://ohsu.eagle-i.net/i/0000012f-7a3b-be79-f561-6e8a80000000
Resource Type
Properties
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Resource Description
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"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."
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Additional Name
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Generating mouse embryo fibroblasts protocol
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Uses
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Cook Medical benchtop embryo incubator
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Uses
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VWR CO2 embryo incubator
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Uses
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Genevac Bio-Cool programmable cryopreservation freezer
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Uses
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BTX ECM 830 electroporator
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Uses
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Electroporation
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Uses
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Establishing cell culture
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Used by
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Transgenic Mouse Model Shared Resource
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Website(s)
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http://www.ohsu.edu/xd/research/research-cores/transgenics/protocols.cfm