Project BriefTools For DNA Diagnostics (October 1994)Diagnostic Laser Desorption Mass Spectrometry Detection of Multiplex Electrophore Tagged DNADevelop a powerful DNA sequencing instrument that combines electrophoretic separation of multiple target DNA segments with specific tags that are rapidly identified with a mass spectrometer. Sponsor: Bruker Daltonics, Inc.19 Fortune DriveBillerica, MA 01821
A four-way collaboration led by Bruker Analytical Systems, Inc., jointly proposes to develop DNA sequencing instrumentation capable of sequencing in one year the equivalent of 10 entire human genomes. The instrument combines sophisticated variations on two standard analytical themes--electrophoretic separation of molecules and mass spectrometric identification of the molecules--into a new analytical system suited for rapidly sequencing DNA samples. A key to the electrophoretic component is the chemical synthesis of several hundred distinct though closely related "electrophores," which link specifically to each member of a library of short DNA probes. These electrophore-tagged probes then hybridize, or bond, to the respective regions of target DNA molecules whose sequences are complementary to that of the probes. Using an existing technique such as PCR (polymerase chain reaction), these tagged probes that hybridize to target molecules will be replicated and then loaded onto electrophoresis gels, where the different probes will spread out and segregate within the gel's matrix. This is where the mass spectrometry comes in. As a laser energizes each segregated spot serially, the electrophore tag at each location feeds into a mass spectrometer that can distinguish its chemical identity and, thereby, the associated DNA sequences. From that data, the sequence of the target DNA can be reconstructed using known protocols.
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