98
DNA
Amplification of the Whole Genome from a Single Cell--Molecular
Staging Incorporated, 300 George Street, 7th Floor, New Haven, CT
06511-6652; 203-772-5000, www.molecularstaging.com
Dr.
Roger Lasken, Principal Investigator,
Dr.
DOE
Grant No. DE-FG02-02ER83511
Amount:
$750,000
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.