Genosensor Matrices for Genome Mapping and Sequencing

This project will use sequencing by hybridization (SBH) to develop a faster, more economicalmethod for sequencing and mapping. Current DNA sequencing procedures almost universally require gelelectrophoresis of DNA fragments and are cumbersome and slow. SBH promises to produce rapidly largeamounts of sequence information due to the parallel nature of the analysis and is well suited forsequencing cDNAs or partial sequencing of amplified DNA to facilitate mapping. Enriched stable isotopeswill be utilized as labels for SBH to demonstrate the basic advantages of such labels and to show themultiplexing feasibility. We have demonstrated that stable isotopes function as both DNA andoligonucleotide labels and that resonance ionization spectroscopy can offer a very fast method for thedetection of surface-bound DNA. During Phase I we will be evaluating two potentially viable detectiontechniques for rapid sequencing and mapping using genosensor matrices: sputter-initiated resonanceionization spectroscopy (SIRIS) and laser atomization RIS (LARIS). Measurements will be made on nylonsurfaces, onto which DNAs have been attached, to identify positively hybridized and unhybridized sitesand to determine the sensitivity, efficiency, background, spatial resolution, and speed and theinterrelationships and trade-offs among them.