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, Perelman School of Medicine, University of Pennsylvania.
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.
RNA purification for CRISPR injection »
Courtesy of Dr. Jorge Henao-Mejia, Department of Pathology and Laboratory Medicine, Perelman School of Medicine, University of Pennsylvania.
Generating Large (>1kb) Genomic Deletions Using CRISPRs»
Morrisey Lab Protocol, by Dan Swarr & Dave Frank.