Purification of BAC DNA for high-efficiency transgenesis

An unresolved bottleneck in bacterial artificial chromosome (BAG) transgenesis is low efficiency generation of founder mice because of suboptimal quality of the manipulated BAG DNA. Using mini-gel electrophoresis and electro-elution that circumvents CsCl(2) centrifugation, column chromatography, and resin purifications, we have used RECOCHIP, a commercially available dialysis cassette for the purification of BAG DNA that generates transgenic founders with up to 80% efficiency.