Novel Two-Dimensional LIF Detector for DNA Sequencing
DESCRIPTION (Applicant's abstract): This Phase I SBIR project introduces a
novel laser-induced fluorescence (LIF) detector that will raise DNA sequencing
to a new level. All commercial sequencers make basecalls from one-dimensional
fluorescence data, viz., intensities at four wavelengths. Fluorescence
lifetime, another basecalling option, is developmental. Our concept uniquely
generates fully two-dimensional data arrays as complete fluorescence decay
profiles are measured at several wavelengths simultaneously. The approach
relies on a high repetition rate pulsed laser, has no moving parts, and uses a
simple photomultiplier tube instead of a CCD camera. Of the many benefits, the
most notable is that the two-dimensional data format offers a multiplicative
improvement in basecalling accuracy. If the probability of an inaccurate
basecall is 5 percent for spectral data and 10 percent for lifetime data, the
combined data reduces the inaccuracy to 0.5 percent. Moreover, the ability to
resolve overlapping peaks and discriminate against background fluorescence
improves dramatically, allowing far longer fragments to be sequenced in much
shorter time. Projected performance by the end of Phase II includes ability to
measure in parallel on 16 capillaries, read length greater than 1000 bases with
better than 99.5 percent basecalling accuracy, and less than 90-minute run
time. No commercial instrument comes close to this combination of speed and
PROPOSED COMMERCIAL APPLICATION:
Automated DNA sequencers have revolutionized genetic research and diagnostic procedures.
Annual sales of $200 million were estimated for automated DNA sequencing equipment in
1999 and the size of the market is expected to increase sharply during the next five years.
The need for an automated DNA sequencer that can provide more accurate basecalling,
longer real lengths, and shorter run times will be met by the proposed research, in which the
first two-dimensional fluorescence detector for sequencing will be developed.
Small Business Information at Submission:
Principal Investigator:Gregory D. Gillispie
DAKOTA TECHNOLOGIES, INC.
2201-A 12TH ST N FARGO, ND 58102
Number of Employees: