Ordering Information Abstracts from Past Meetings 152. Genome Shotgun Mapping Anantharaman, T, Apodaca, J., Aston, C., Clarke, V., Gebauer, D., Delobette, S., Dimalanta, E., Edington, J., Giacalone, J., Gibaja, V., Huff, E., Jing, J., Lai, Z., Lin, J., Limm, A., Mishra, B., Ni, L., Paxia, S., Qi, R., Ramanathan, A., Skiadis, Y., Vafai, J., Wang, W., Schwartz, D.C. University of Wisconsin - Madison Optical Mapping is a single molecule approach for the rapid production of ordered restriction maps from single DNA molecules. Fluorescence microscopy is used to image individual DNA molecules bound to derivatized glass surfaces, and cleaved by restriction enzymes.
Fragments retain their original order, and cut sites are flagged by small, visible gaps.
We mapped these entire microbial genomes using megabased-sized genomic DNA molecules. Because large fragments of randomly sheared DNA are mapped with high cutting efficiency, many overlapping restriction site landmarks allow contigs to be assembled and a shotgun mapping strategy can be employed. Because ensembles of single molecules are analyzed, small amounts of starting material are required enabling mapping of microorganisms which are problematic to culture. Whole genome maps firstly, enable the size of the genome to be accurately determined, an important prelude to any sequencing endeavor and secondly, provide an in situ picture of the architecture of the entire genome, revealing the number of chromosomes, extrachromosomal elements etc. Populations can be potentially be characterized by comparing maps from different strains.