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Multiplexed DNA sequencing and diagnostics by hybridization with enriched stable isotope labels
H F Arlinghaus1, M N Kwoka, X Q Guo
1Atom Sciences, Inc., Oak Ridge, Tennessee 37830, USA.
Analytical Chemistry
|April 15, 1997
Summary
A new DNA sequencing method uses stable isotopes and time-of-flight resonance ionization mass spectrometry (TOF-RIMS) for rapid analysis. This technique enables simultaneous detection of multiple DNA probes, enhancing diagnostic capabilities.
Area of Science:
- Biochemistry
- Analytical Chemistry
- Molecular Biology
Background:
- Sequencing by hybridization (SBH) is a method for DNA analysis.
- Current SBH methods face challenges in multiplexing and detection sensitivity.
Purpose of the Study:
- To develop a rapid DNA diagnostic and sequencing system.
- To utilize stable isotope labeling and TOF-RIMS for enhanced SBH analysis.
Main Methods:
- DNA immobilized on nylon membranes.
- Hybridization with stable isotope-labeled oligonucleotide probes.
- Detection using time-of-flight resonance ionization mass spectrometry (TOF-RIMS).
Main Results:
- Simultaneous hybridization and detection of multiple probes achieved.
- High selectivity and sensitivity demonstrated with isotopically enriched tin labels.
- Discrimination between complementary and noncomplementary sites exceeded 100:1.
Conclusions:
- The new SBH method offers a versatile and extensive multiplexing capability.
- Stable isotopic labels combined with TOF-RIMS provide efficient DNA detection.
- This approach significantly advances DNA sequencing and diagnostics.
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