[Electron
Microscopy] [Flow Cytometry] [Histology]
[Imagecore] [Nucleotide
Sequencing]
[Peptide Core] [Transgenic Core] [VTPB
Central Stores] [VTPB Grading
Facility]
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 Kier (1-979-845-3365).
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 A&M University.
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 males
- Embryo derivation for the elimination
of specific mouse pathogens
Core Equipment:
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 capabilities.
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 vibration.
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 observation.
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.
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