Where do I start?
So you want to make a transgenic or knockout line? Here's what you'll need to have before we can begin work on your project:
- Approved animal protocol in place
- DNA prepared using protocols provided by Core
- Plan in place for screening ES cells (for KO)
- Assigned animal space as destination for mice
- Plan in place for screening pups
- Trained personnel to breed mice, cut tails, and screen DNA
- UCSD index number or an off-campus purchase order
If you require a support letter from the Transgenic Core for your grant submissions, please contact MellonAdmin@ucsd.edu.
Forms and protocols
The following forms and protocols require Adobe Acrobat Reader to be installed on your computer's hard drive in order to be downloaded for viewing. If you do not have Adobe Acrobat Reader, click here.
UCSD Transgenic Core Request For Services - This form MUST be filled out before beginning any work with the Core.
DNA Preparation for Pronuclear Injection - Contact Core personnel for current Pronuclear Injection protocol.
DNA Preparation for Gene Targeting Construct - Here is the final preparation protocol you must use before we transfect your Gene Targeting Construct into ES Cells.
Genomic DNA Extraction from Mouse Tissue - Now that we've given you potential transgenic pups, here is an efficient way to extract DNA from tail fragments for Southern Blot and PCR analysis.
Genomic DNA Preparation from Mouse Tissue - PCR only - This is a phenol-free DNA preparation that can be used with 1-2 mm of mouse tissue for PCR analysis only.
Genomic DNA Extraction from 96-well Plates - Use this method to extract DNA for Southern Blot and PCR analysis of targeted ES cell clones from the duplicate 96-well plates.
Cell Preparation Protocol for Blastocyst Injection - Do you have your own homologous recombinants that you want us to inject into Blastocysts? Use this protocol to prepare your ES cells.