The objective of this program is to develop a technique that can elucidate structural information from complex nucleic acids. Ideally, any technique employed for nucleic acid analysis should yield not only structural details which will reveal the nucleotide sequence, but will also provide the end user with three dimensional conformational information. For example, the ability to analyze variations in the three dimensional structure of the DNA double helix can lead to an improved understanding of the details of the transfer of genetic traits. We propose to elucidate structural and conformational information by probing the chiral centers within the nucleic acids via the rapidly maturing spectroscopic technique of Raman Optical Activity (ROA). Efficient utilization of the traditional Raman spectrum as well as the ROA spectrum will enable the end user to determine structural and conformational information from nucleic acid samples. The Phase I objective is to determine the feasibility of rapid structural and conformational identification of nucleosides and nucleotides, the nucleic acid building blocks, with ROA spectroscopy. During the Phase I program we will attempt to establish the sensitivity levels at which ROA will be capable of rapid detection of nucleic acids and will collect representative spectra from a nucleic acid. During the Phase II program a more detailed analysis of several nucleic acids will be pursued as well as detailed spectral studies of small (2-4 nucleotide units) oligonucleotides in order to correlate certain ROA spectral features with specified nucleotide sequences and conformations. During the Phase II program a prototype ROA sampling accessory, which can be implemented into any commercial Raman system, will be developed.
