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Single Molecule Detection in Flowing Sample Streams As An Approach to DNA Sequencing

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Abstract

We are exploring a technique which has the potential to sequence large fragments of DNA at a rate of hundreds of bases per second. Our technique is based upon a projected ability to detect single chromophores by laser-induced fluorescence in flowing sample streams.1 The technique involves: (1) labeling the nucleotides with base specific tags suitable for fluorescence detection, (2) selecting a desired fragment of DNA, (3) suspending the single DNA fragment in a flowing sample stream, (4) sequentially cleaving labeled bases from the free end of the DNA fragment using an exonuclease, and (5) detecting and identifying the cleaved, labeled bases as they flow through a focused laser beam.2

© 1990 Optical Society of America

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