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Current status of DNA sequencing by single molecule detection

Research output: Chapter in Book/Report/Conference proceedingConference contribution

14 Scopus citations

Abstract

Our current experiments further the development of a laser-based technique capable of sequencing an individual strand of DNA. We report the detection and identification of fluorescently labeled nucleotides enzymatically cleaved from DNA strands suspended in flow. We used fluorescence lifetime, fluorescence intensity, or a correlated measure of the intensity and lifetime to identify each individual tagged base traversing the detection region with high accuracy. DNA strands containing a single tetramethylrhodamine labeled uracil and/or a single Rhodamine 6G labeled cytosine were attached to polystyrene microspheres. An optical trap was used to capture and hold a single DNA-laden microsphere nominally 20 microns upstream of the detection region of an ultra-sensitive flow cytometer. The addition of an exonuclease cleaved bases from the 3' end of the fluorescently labeled strand. The cleaved, labeled nucleotides were carried by the flow downstream and detected and identified one-at-a-time with high efficiency by laser-induced fluorescence.

Original languageEnglish
Title of host publicationProceedings of SPIE - The International Society for Optical Engineering
PublisherUnknown Publisher
Pages355-366
Number of pages12
ISBN (Print)0819430722
StatePublished - Jan 1 1999
EventProceedings of the 1999 Advances in Fluorescence Sensing Technology -
Duration: Jan 1 1999 → …

Publication series

NameProceedings of SPIE - The International Society for Optical Engineering
Volume3602
ISSN (Print)0277-786X

Conference

ConferenceProceedings of the 1999 Advances in Fluorescence Sensing Technology
Period01/1/99 → …

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