Homologous recombination Homologous recombination EXon 2( Upper half) green= homologous DNA Cut and ligated Cut and ligated HS Neo r( lower half) HSVtk complete double strand DNA Random Integration Neo r Cut and inserted
Homologous Recombination Homologous recombination Random Integration Cut and ligated Cut and ligated Cut and inserted Exon 2 ( Upper half) Neo r ( lower half ) HSVtk complete double strand DNA Neo r HSVtk green= homologous DNA HSVtk
ES= embryonic stem cells are totipotent, they can develop into any tissue Culture on feeder cells with selective medium Ganciclovir and neomycin nner cell mass Culture of Blastocyst embryonic stem (ES)cells Electroporation Trypsinise and culture Uptake of knockout vector DNA via a on feeder cells short electric pulse. Efficiency: about 10%-30% of the cells are transfected
Culture on feeder cells with selective medium Ganciclovir and Neomycin Trypsinise and culture on feeder cells Uptake of knockout vector DNA via a short electric pulse. Efficiency: about 10%-30% of the cells are transfected ES = embryonic stem cells are totipotent , they can develop into any tissue