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Transgenics
Gene Targeting
Chimeric Mice
Rederivation
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New ES Cells
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Frozen Embryo Transfer
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General Mouse Protocols:
Breeding, Care And Maintenance Of Mice
Plasmid Transgene DNA Purification For Pronuclear Injection
Purification Of BAC DNA For BAC Transgenics
Detecting Transgenes by PCR and Southern Blot
Preparing Earpunch DNA For PCR Genotyping
Preparing Tail Or Toe DNA For Southern Blot Analysis
Performing Gavage on Mice
Fostering Mice
David LePage's Gene Targeting Protocols:
•
LePage DF, Conlon RA. (2006) Animal models for disease: knockout, knock-in, and conditional mutant mice.
Methods Mol Med
129:
41-67. (.pdf)
•
Dave's Bench Gene Targeting Protocols in pdf form
Growing ES Cells
Passaging From 35 mm Dishes
Freezing From 35 mm Dishes
Preparing Vector DNA
Electroporation
Picking Lines
Tryplating
Freezing From 96-Well Plates
Extracting Genomic DNA From 96-Well Plates
Cutting Genomic DNA In 96-Well Plates
Southerns
Thawing 96-Well Plates
Extracting Genomic DNA From 24-Well Plates Or Larger
Counting ES Cell Chromosomes
Counting ES Cell Chromosomes: Examples
Preparing ES Cells For Blastocyst Injection
David LePage's Protocols for Making New ES Cell Lines:
•
pdf file
References
Schedule
Initial Culture Conditions
Embryo Collection
Disaggregation
Passaging
Guidelines for Growing New Lines