We propose to develop prepolymer-based gelling chemistries that are compatible with DEL sequencing and protein gel electrophoresis to make gels at concentrations much lower than the usual acrylamide/bis gels. The need for increasing throughput of DEL sequencing is well known. If the readout rate can be increased to more than 1200 bases per reaction (b/rxn), the reduction in overlap sequencing and sequence assembly steps could lead to an increase in the net throughput rate of several-fold. If gels can be run at as low as 3%T (total concentration), then the base readout rate of a given technique is expected to be almost doubled. Thus readout rates of over 1500 in automated sequencing and over 1000b/rxn in manual techniques may be attainable. Commercial prepolymers can set strong enough gels at 3%T, but their chemistries are incompatible with DNA sequencing. Several different prepolymer gel chemistries will be evaluated for: Their ability to set gels as strong as 5%T acrylamide/bis gels at inch lower %T; and their ability to attain single base DNA resolution in sequencing. These prepolymer gels will also be useful for resolving clinically important large proteins, and protein-nucleic acid assemblies.
Successful project will result in novel prepolymer gels for DEL sequencing and for very large protein electrophoresis. The potential market is over $100 million. Prepolymer technology will later enable precoated ultrathin gels for appropriately designed clinical instruments for rapid, definitive, DNA-oriented tests for infections, cancers, and genetic markers, This market is expected to be several billion dollars.
