Es syllabus 2012
The University of Michigan 2570 MSRB II, SPC 5674 1150 W. Medical Center Drive Transgenic Animal Model Core Ann Arbor, MI 48109-0674 TEL: (734) 647-2910 FAX: (734) 936-2622 [email protected] Linda C. Samuelson, Ph.D. Thomas L. Saunders, Ph.D. Mouse Embryonic Stem (ES) Cell Training The purpose of the class is to provide training in all aspects of ES cell culture manipulation and to provide the scientific background needed to make a gene targeted (gene knockout) mouse. You will both methods and the principles behind the methods. The Mouse Embryonic Stem Cell Training Course is designed to instruct researchers in the art of pluripotent mouse ES cell culture and the methodologies of gene targeting. The training includes daily laboratory sessions on culture techniques. Daily seminars cover key papers in the field of gene targeting, conditional gene knockouts, and genetic background (see bibliography). Trainees are expected to read papers, participate in discussions, and present at least one seminar. Special topics specific to trainee research are welcome. The course will take place in room 2578 Medical Science Research Building II, and will last two weeks. In order to provide as much hands-on experience as possible, ES cell training class size is limited to four or fewer trainees. A. Purpose: To instruct personnel in the art of ES cell culture and gene targeting protocols. B. Training Overview: Discussion and lab experience in ES cell culture methods and techniques. This includes: setting up a laboratory for stem cell work, evaluation of fetal bovine serum lots for ES cell culture, mycoplasma testing, media preparation, train eye for stem cell differentiation, normal ES culture, preparation of mouse embryonic fibroblast (MEF) for co-culture, freezing and thawing of ES cells, ES cell electroporation with DNA, ES cell clone picking, cryopreservation and recovery of ES cell clones in 96-well plates, DNA preparation from cells cultured on 96 well plates, preparation of chromosome spreads from ES cells and chromosome counting. C. Preparation: C1. Highly Recommended: Gene Targeting: A Practical Approach. 2000. Joyner AL, ed. Oxford University Press; ISBN: 019963792X. Read the entire book. Or "Advanced Protocols for Animal Transgenesis: An ISTT Manual." 2011. Pease S and Saunders TL, eds. Springer. ISBN 364220791X. Read Chapters 3, 4, 14, and 15. C2. Optional: Read Chapters 8, 9, and 10 in "Manipulating the Mouse Embryo: A Laboratory Manual." 3rd ed. 2003. Nagy, Gertsenstein, Vintersten, and Behringer. Cold Spring Harbor Laboratory Press. pp. 359-451. C3. Optional: read Chapters 49 through 53 in "Guide to Techniques in Mouse Development." 1993. Wassarman and DePamphilis. Methods in Enzymology, Volume 225:803-900. C4. Novices in tissue culture are expected to read "Culture of Animal Cells: A Manual of Basic Technique and Specialized Applications." 6th ed. 2010. Freshney. Wiley-Blackwell. We expect that trainees will be familiar with basic tissue culture procedures.