The Department of Pathobiology has established a core facility
for the production of transgenic and gene targeted mice. The core
is composed of three principal elements:
- Molecular analysis and development of gene constructs for
pronuclear injection and gene targeting.
- Production of transgenic mice by pronuclear injection
(overexpression) and blastocyst injection (gene targeting).
- Detection, analysis and breeding of transgenic mice.
This core facility is supervised by Dr. Ann
Further core facilities are available in the department for the
morphometric analysis of transgenic animals and their tissues. The
animals are maintained in a specific pathogen free (SPF) barrier at
the Laboratory Animal Resources and Research Facility at Texas
Dr. Kier will advise and assist in the development of protocols
utilizing the following methodologies:
- Embryonic stem (ES) cell library screening - mouse
- ES cell culture
- Electroporation of ES cells
- DNA screening by Southern analysis and PCR
- Superovulation and embryo collection and transfer
- Pronuclear microinjection
- Blastocyst microinjection
- Analysis of sperm by PCR for the evaluation of transgenic
- Embryo derivation for the elimination of specific mouse
Inverted microscope: A Nikon Diaphot
300 equipped for brightfield, phase contrast, Nomarski differential
interference contrast and epi-fluorescence observation. The
microscope has both 35 mm still photography and video
Micromanipulators: A pair of
Narshige MO-188 and MN-188 hydraulic micromanipulators are mounted
on the Nikon Diaphot for the holding and injection of embryos.
Inverted microscope table: A gimbal
gas piston isolator for vertical and horizontal isolation of
Pipet Puller: A Stoelting APP-1
microprocessor controlled horizontal pipet puller. This pipet
puller is used for the production of injection pipets which can be
accurately and repeatedly reproduced.
Microscope: A Nikon TMS inverted
microscope equipped for brightfield and phase contrast
Biologic Safety Cabinet: A Nuaire
model NU425-400 HEPA filter vertical laminar flow hood is used for
tissue culture procedures.
CO2 incubator: A Nuaire model
NU-2500 CO2 incubator which is equipped with infrared CO2
monitoring, HEPA filtered air and humidity monitoring, is used for
tissue and embryo culture procedures.