DNA Amplification of the Whole Genome from a Single Cell

70555S02-II DNA sequencing is currently not possible when starting from single bacterial cells because of the limited amount of DNA present. Typical medical and biological samples contain a heterogeneous mixture of species; therefore, obtaining a sufficient amount of DNA requires a time-consuming and labor-intensive approach involving the isolation and culture of bacterial cells. Since most bacteria are simply unculturable, the vast number of these highly valuable microorganisms remains unstudied. This project will develop a kit to amplify the whole bacterial genome from a single cell, allowing DNA sequencing that was previously not possible in many research projects. In Phase I, a proof-of-concept was obtained for whole-genome amplification from a single Escherichia coli cell. In Phase II, the DNA amplification technique will be extended to other bacteria that represent the diversity of commercially and medically important species encountered in natural field samples. Methods will be refined for cell sorting, cell lysis, and DNA amplification. Commercial Applications and Other Benefits as described by awardee: The ability to sequence DNA from a single bacterial cell should greatly accelerate progress in many medical and basic research areas. For example, it should be possible to accelerate the preservation of genetic archives at a time of vanishing habitats such as rain forests and coral reefs. DNA sequencing from single cells also offers an opportunity to develop new and sensitive methods to detect and trace the location of origin of microbes for medical and biodefense applications.