Seasoned practitioners from many leading laboratories describe their best readily reproducible screening strategies for isolating useful clones. These techniques have been optimized for sensitivity, high throughput, and robustness, and are of proven utility for directed evolution purposes. The assays presented use a variety of techniques, including genetic complementation, microtiter plates, solid-phase screens with colorimetric substrates, and flow cytometric screens. An accompanying volume, Directed Evolution Library Creation: Methods and Protocols, describes readily reproducible methods for the creation of mutated DNA molecules and DNA libraries.... of the pCWOri HRP vector works well with many E. coli cell strains. However, for ... For mL-scale cultures, manual pipetting can be done with the tip of the pipet completely immersed in the liquid, such that no air bubbles are introduced.
|Title||:||Directed Enzyme Evolution|
|Author||:||Frances Hamilton Arnold, George Georgiou|
|Publisher||:||Springer Science & Business Media - 2003-01-01|