Protocol
Protocol
protocol
protocol
Generating MEFs protocol
http://www.ohsu.edu/xd/research/research-cores/transgenics/protocols.cfm
Website(s)
has URL
has_url
"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."
Resource Description
resource_description
Generating mouse embryo fibroblasts protocol
Additional Name
eagle-i_synonym
Uses
Electroporation
specifies the use of
electroporation
specifies_the_use_of
electroporation
Establishing cell culture
establishing cell culture
establishing cell culture
Cook Medical benchtop embryo incubator
VWR CO2 embryo incubator
Genevac Bio-Cool programmable cryopreservation freezer
BTX ECM 830 electroporator
Used by
used by
used_by
Transgenic Mouse Model Shared Resource
2011-04-21T17:47:06.272-05:00
nvasilevsky (Nicole Vasilevsky)
2011-04-21T17:49:52.820-05:00
workflow state
Published
nvasilevsky (Nicole Vasilevsky)
Document
document
document
information content entity
information content entity
continuant
entity
generically dependent continuant