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Training
Transgenic Animal Model
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Embryonic Stem Cell Culture Training
Participants will learn all techniques necessary to produce and identify
embryonic stem (ES) cells that have undergone homologous recombination with a
targeting vector. Research papers providing the experimental background
underpinning methods used to engineer gene targeted mice will be presented and
discussed daily. Laboratory training includes setting up a lab for ES cell
culture, media and reagent preparation, normal ES cell culture, preparation of
feeder cells, freezing and thawing of ES cells, identification of differentiated
and undifferentiated ES cell morphologies, ES cell electroporation, picking ES
cell clones, DNA preparation from ES cell clones in 96 well plates, and counting
chromosomes in ES cell spreads. Download a
Syllabus.
Pronuclear Microinjection Training
Participants will learn all techniques necessary to produce transgenic mice.
This includes superovulation, glass microinstrument fabrication, egg collection,
identification of pseudopregnant females, surgical transfer of eggs to
pseudopregnant recipients, and microinjection of DNA into the pronuclei of
fertilized mouse eggs. A majority of trainees produce transgenic mice during
training. Download a Syllabus.
Rat transgenic production training is also available. Download a Syllabus.
Blastocyst Microinjection
Participants will learn all techniques necessary to produce embryonic stem
cell-mouse chimeras. This includes superovulation, glass microinstrument
fabrication, blastocyst collection, identification of pseudopregnant females,
surgical transfer of blastocysts to pseudopregnant recipients, and
microinjection of ES cells into the blastocoels of C57BL/6 blastocysts. A
minority of trainees produce chimeric mice during training.
Mouse Embryo Transfer
Participants will learn all techniques necessary to transfer mouse embryos to
the reproductive tract of recipients. This includes superovulation, isolation of
pathogen free fertilized eggs from infected stocks, transfer pipette
fabrication, identification of pseudopregnant females, surgical transfer of eggs
to pseudopregnant recipients. All trainees produce offspring from transferred
eggs during training.
Consult on design and construction of transgenes and targeting vectors; identification of transgenic founders; and breeding.
