Protocols

These guidelines have been collected and compiled from the notes of investigators using the facility, as well as from the experiences of the facility's technical personnel. The protocols are also available for download in the "Manual & Forms" section.

• DNA Purification
  Two recommended procedures are provided here for your convenience. Each has been thoroughly tested and are equally effective in producing transgenic mice using DNA constructs.
  
  
• DNA Extraction
  Protocol for isolating high-molecular-weight DNA from mouse tails (from Manipulating The Mouse Embryo; Hogan et al., Cold Spring Harbor, 1986).
  
  
• Preparation of BAC DNA for Pronuclear Injection
  Courtesy of Dr. Douglas Epstein, Department of Genetics, University of Pennsylvania School of Medicine.
  
  
• Preparation of HNF3g YAC DNA for Microinjection
  Protocol for the correct preparation of YAC DNA as well as for the large scale preparation of yeast plugs and various related solutions are provided here.
  
  
• Preparation of ES Cells
  Procedure for preparation of a sample of ES cells for injection into blastocysts.
  
  
• Karyotyping ES cells
  A protocol that describes how to prepare chromosome spreads from ES cells.
  
  
• Animal Husbandry
  General guidelines and suggestions for basic mouse husbandry. Please contact the TCMF at 215/573-3023 if you have additional questions.