The project proposes a strategy aimed at increasing by an important factor the output of electrophoretic sequencing, mapping and DNA-probe genetic diagnosis, by consistently integrating innovations in different fields in which the proposers have specific competences.
The most important of those innovations, each of which represents a valuable intermediate target, are:
-New improved permanent and non-permanent sieving matrices for sequencing and duplex DNA separation
-Design of an automated capillary array electrophoresis apparatus able to load from Microtiter plates, and to perform constant-field and pulsed-fields electrophoresis.
-Alternative strategies using neutral molecules or particles bound to DNA to increase band separation.
-Improved injection protoeols for capillary electrophoresis to avoid prepurifieation steps and increase automation
-Comprehensive predictive models to accelerate the optimization of sequencing and DNA assay using pulsed-fields and/or polymer solutions. The project corresponds to area III.5 of the call for proposals, and more partieularly to the tasks: "development of new or improved sequencing approaches..." and "application of new methods of DNA probe diagnosis and their automation..." The whole projeet will be driven by fundamental studies including:
-Theoretical modelling and computer simulations
-Extensive studies of molecular migration mechanisms and of collective behavior by fluorescence microscopy.
This approach, which requires multiple acute competences but no dramatic and hypothetical technlogieal breakthrough, would provide a rather secure and short route to accelerated sequencing and mapping, and automatised diagnosis.
The chances of success of the project are maximised by exploring several parallel solutions for the most critical aspects (e.g.
sieving matrices, increase of resolution).
Funding SchemeCSC - Cost-sharing contracts
20090 Milano - Segrate