The documents listed below are detailed protocols for the new and experienced ES cell/mouse user alike. These protocols have been tested and optimized repeatedly, and should provide an efficient means to reproducible results. Some protocols include MIT-specific information, though all should be applicable in general.
If you have a protocol that would be useful for other users, please contact Aurora and Noranne (firstname.lastname@example.org) and we'd be delighted to add it to this site. Also, if you have any questions about any part of a protocol, please let Aurora and Noranne know. We strive to provide clear, easy-to-use protocols to reduce the hassle and mystery involved with ES cell culture, gene targeting and mouse models. Your feedback helps us reach that goal.
- Beginners' Guide to Gene Targeting
- Targeting Construct Linearization
- Making Hot Probes For Southerns
- Southern Blots with Alkaline Transfer
- Southern Blots with Neutral Transfer
- Generating a Transgenic Mouse
- Preparing Cas9 mRNA and CRISPR gRNA for Pronuclear Injection (please email us for a copy)
- Preparing ES cells for Blastocyst Injection
- Beginner's Guide to Mouse Management
- Isolating and Culturing MEFs or Feeders
- Blastocyst Harvest and Culture
- Karyotyping a Cell Line
There are more protocols and reagent information for ES cell culture to be found under Useful Links.