NSTI Nanotech 2009

(CANCELED) Massively Parallel Affinity Selection with Complex Selectors

G.P. Smith, W.D. Thomas
University of Missouri, US

Keywords: phage, diversity, in vivo selection, complex selectors, deep sequencing

Abstract:

In a “massively parallel” (MP) affinity selection, the selector is a complex mixture of hundreds, thousands or millions of target molecules. MP affinity selection inevitably entails some degree of compromise between two countervailing desirables. On the one hand, it is usually highly desirable to preserve the diversity of the output phage so that it can be thoroughly characterized; this is accomplished by reducing stringency—usually by reducing the number of rounds of selection. On the one hand, it is also highly desirable to reduce the complexity of the output phage population sufficiently to characterize that population adequately; this is accomplished by increasing stringency. Ordinarily, even if an optimum stringency could be defined, it is entirely unknown in advance how to achieve. “Deep sequencing” would largely eliminate the need for compromise. Deep sequencing is a generic name for one of the new generation of technologies that is able to sequence tens of millions of clones in a single run. It would allow an output phage population with up to a million individual clones to be thoroughly characterized after only one or two rounds of affinity selection.
 
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