Purification of BAC DNA for high-efficiency transgenesis
Rajendra K Gangalum,
Zhe Jing,
Yoshiko Nagaoka,
Meisheng Jiang,
Suraj P. Bhat
Affiliations
Rajendra K Gangalum
1Jules Stein Eye Institute, Los Angeles, CA, USA
Zhe Jing
1Jules Stein Eye Institute, Los Angeles, CA, USA
Yoshiko Nagaoka
3Department of Molecular and Medical Pharmacology, and Jonsson Cancer Center, Geffen School of Medicine at UCLA, University of California, Los Angeles, CA, USA
Meisheng Jiang
3Department of Molecular and Medical Pharmacology, and Jonsson Cancer Center, Geffen School of Medicine at UCLA, University of California, Los Angeles, CA, USA
An unresolved bottleneck in bacterial artificial chromosome (BAC) transgenesis is low efficiency generation of founder mice because of suboptimal quality of the manipulated BAC DNA. Using mini-gel electrophoresis and electro-elution that circumvents CsCl2 centrifugation, column chromatography, and resin purifications, we have used RECOCHIP, a commercially available dialysis cassette for the purification of BAC DNA that generates transgenic founders with up to 80% efficiency.
