DNA Base Calling using Heterodyne Fluorescence Detection
DESCRIPTION (provided by applicant): Currently available high throughput DNA
sequencing instruments achieve base calling by wavelength resolved,
fluorescence detection of dye labeled fragments using a CCD camera.
Identification of labels using fluorescence lifetimes is also possible. Because
cost effective, high speed, multichannel detectors are not available, adapting
lifetime detection to high throughput sequencing would seem to require
timesharing of a single element detection system between multiple
electrophoresis channels. Southwest Sciences has developed a heterodyne
fluorescence lifetime method that is readily scalable to multi-element
detection. This approach uses simple, relatively low cost detection electronics
and a diode laser excitation source and will perform competitively with current
instrumentation at lower cost.
In Phase I, we successfully demonstrated the necessary time resolution,
sensitivity, and speed needed for DNA sequencing applications using a single
element detector and aqueous dye solutions. The Phase II effort will focus on
adapting this technique to capillary array sequencing. A single-capillary
instrument with an optimized heterodyne detector will be used to investigate
the base calling accuracy of this method in four-dye, single lane sequencing.
Analysis methods will be developed to call bases from the lifetime data.
Finally, a 16 element heterodyne detector, suitable for capillary arrays, will
be constructed and evaluated.
PROPOSED COMMERCIAL APPLICATION:
Automated DNA sequencing has application to genetic testing for medical diagnostics, agricultural and pharmaceutical applications. Our heterodyne approach to DNA base calling will significantly lower the cost of automated DNA sequencing instrumentation.
Small Business Information at Submission:
Principal Investigator:Kristen A. Peterson
SOUTHWEST SCIENCES, INC.
1570 PACHECO ST, STE E-11 SANTA FE, NM 87505
Number of Employees: