Purification of BAC DNA for high-efficiency transgenesis. (November 2011)

• Record Type: Journal Article
• Authors: Gangalum, Rajendra K; Jing, Zhe; Nagaoka, Yoshiko; Jiang, Meisheng; Bhat, Suraj P.
• Is Part Of: Biotechniques. Volume 51:Number 5(2011)
• Page Start: 335
• Page End: 337
• Publication Date: 2011-11
• Subjects: BAC DNA -- transgenesis -- RECOCHIP -- Electro-elution; Biology, Experimental -- Periodicals; Molecular biology -- Periodicals; Medical technology -- Periodicals; Biology, Experimental; Medical technology; Molecular biology; Clinical Laboratory Techniques -- Periodicals; Research -- Periodicals; Medical Laboratory Science -- Periodicals; Periodicals; Electronic journals; 570
• DOI: 10.2144/000113763
• Languages: English
• ISSNs: 0736-6205

Abstract

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.