Transgenic Core Facility

EmailEmail    |   Bookmark Page Bookmark  |   RSS Feeds RSS  |   Print Page Print  

Resources


Mice

The Core maintains colonies of C57Bl/6J Stud males for oocyte and blastocyst production; vasectomized males, foster females, neoR-mice for production of neomycin-resistant feeder cells, as well as wild type mice of several defined genetic backgrounds.

ES cells

A number of different germline-competent ES cell lines are available as frozen stocks.

Feeder cells

Frozen stocks of neomycin-resistant primary embryonic fibroblast feeder cells for ES culture.

DNA constructs

DNA vectors containing modules for the construction of gene targeting vectors are available by request.

Genomic DNA

The core can provide control DNA from ES cells and mouse tails that may be used to establish a screening procedure capable of detecting single copy genes.


Equipment

The Core has specialized equipment that can upon request be made available to Investigators. This includes a microinjection station, equipment for tetraploid embryo aggregation, a laminar flow workstation with a surgical microscope located in the transgenic animal barrier facility, teaching stereoscope for microsurgery, dissecting microscope with color video capture, fully automated needle puller, a microforge for production of customized needles/pipettes, and a controlled rate freezer for cryopreservation of cells and embryos. Access to this equipment depends on utilization by the Core and must be pre-arranged with the staff.

 

Microinjection
Station
Dissecting Microscope
w/color video capture
Microforge
for production
of customized needles/pipettes
Controlled Rate
Freezer
for
cryopreservation
of cells and embryos
Gene Pulser
       
 Cell Fusion
Controller for
tetraploid embryo aggregation
 Needle Puller
Fully automated
 Teaching Stereoscope
for microsurgery
 Dissection Video Capture  Stereo Zoom Microscope

Training

The Core can arrange training sessions for individuals to acquire hands-on skills and expertise in the production of transgenic mice, embryonic stem cell culture, and production of ES cell chimeras. The training will enable individuals to successfully conduct these tasks on their own. Specialized training equipment is available.

Training for oocyte and blastocyst injections requires approximately 60 hours over 5-6 weeks. Topics include mouse husbandry, superovulation, harvesting of blastocysts and fertilized eggs, microinjection/blastocyst injection, and transfer of injected eggs or blastocysts into pseudopregnant foster mice.

The ES cell course teaches all the necessary skills to successfully conduct a gene targeting experiment. Topics include preparation of mouse embryonic fibroblast feeder cells, routine culture of ES cells, evaluation of ES cell morphology and ploidy, electroporation, selection and picking of ES cell clones, freezing and DNA preparation. The time frame for this hands-on course is about 5-6 weeks and includes an entire targeting experiment.

We encourage laboratories planning to utilize these techniques on a frequent basis to take advantage of this opportunity. A fee is levied to cover materials, mice, and staff time. Please contact the Core Director to arrange a training session.

webmaster@mcw.edu
© 2014 Medical College of Wisconsin
Page Updated 04/17/2014