High Throughput BAC DNA Isolation for Physical Map Construction of Sorghum (Sorghum bicolor)
- Additional Document Info
- View All
With the aim of constructing a physical map of sorghum, we developed a rapid, high throughput approach for isolating BAC DNA suitable for restriction endonuclease digestion fingerprinting, PCR-based STS-content mapping, and BAC-end sequencing. The system utilizes a programmable 96 channel liquid handling system and associated accessories that permit bacterial cultivation and DNA isolation in 96-well plate format. This protocol details culture conditions that optimize bacterial growth in deep-well plates and criteria for BAC DNA isolation to obtain high yields of quality BAC DNA. The system is robust, accurate, and ralatively cost-effective. The BAC DNA isolation system has been tested during efforts to construct a physical map of sorghum.
author list (cited authors)
Klein, R. R., Morishige, D. T., Klein, P. E., Dong, J., & Mullet, J. E.
complete list of authors
Klein, Robert R||Morishige, Daryl T||Klein, Patricia E||Dong, Jianmin||Mullet, John E