Klaus Dörre, Susanne Brakmann, Michael Brinkmeier, Kyung-Tae Han, Katja Riebeseel, Petra Schwille, Jens Stephan, Timm Wetzel, Markus Lapczyna, Michael Stuke, Raoul Bader, Michael Hinz, Hartmut Seliger, Johan Holm, Manfred Eigen, Rudolf Rigler
{"title":"Techniques for single molecule sequencing","authors":"Klaus Dörre, Susanne Brakmann, Michael Brinkmeier, Kyung-Tae Han, Katja Riebeseel, Petra Schwille, Jens Stephan, Timm Wetzel, Markus Lapczyna, Michael Stuke, Raoul Bader, Michael Hinz, Hartmut Seliger, Johan Holm, Manfred Eigen, Rudolf Rigler","doi":"10.1002/1361-6374(199709)5:3<139::AID-BIO8>3.0.CO;2-%23","DOIUrl":null,"url":null,"abstract":"<p>A method is described that demonstrates a new technique for rapid and high-throughput single molecule sequencing. This sequencing technique is based on the successive enzymatic degradation of fluorescently labeled single DNA molecules, and the detection and identification of the released monomer molecules according to their sequential order in a microstructured channel.</p><p>The detection technique is evolved from confocal fluorescence microscopy, with two different laser sources to excite the individual mononucleotides that are either labeled with tetramethylrhodamine (TMR) or Cyanine5 (Cy5). The handling of DNA which is immobilized on carrier beads, and the detection of the cleaved monomers is performed in optically transparent and biochemically inert microstructures (glass or PMMA) with detection channels of 7 μ × 10 μm.</p><p>The projected rate of sequencing is ≈100 bases min<sup>−1</sup>, dependent solely on the rate of the enzymatic DNA cleavage.</p>","PeriodicalId":100176,"journal":{"name":"Bioimaging","volume":"5 3","pages":"139-152"},"PeriodicalIF":0.0000,"publicationDate":"2001-05-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/1361-6374%28199709%295%3A3%3C139%3A%3AAID-BIO8%3E3.0.CO%3B2-%23","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Bioimaging","FirstCategoryId":"1085","ListUrlMain":"https://onlinelibrary.wiley.com/doi/10.1002/1361-6374%28199709%295%3A3%3C139%3A%3AAID-BIO8%3E3.0.CO%3B2-%23","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"","JCRName":"","Score":null,"Total":0}
引用次数: 0
Abstract
A method is described that demonstrates a new technique for rapid and high-throughput single molecule sequencing. This sequencing technique is based on the successive enzymatic degradation of fluorescently labeled single DNA molecules, and the detection and identification of the released monomer molecules according to their sequential order in a microstructured channel.
The detection technique is evolved from confocal fluorescence microscopy, with two different laser sources to excite the individual mononucleotides that are either labeled with tetramethylrhodamine (TMR) or Cyanine5 (Cy5). The handling of DNA which is immobilized on carrier beads, and the detection of the cleaved monomers is performed in optically transparent and biochemically inert microstructures (glass or PMMA) with detection channels of 7 μ × 10 μm.
The projected rate of sequencing is ≈100 bases min−1, dependent solely on the rate of the enzymatic DNA cleavage.
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