DOI: 10.1002/elps.11501301125. High‐throughput automated DNA sequencing facility with fluorescent labels at the European Molecular Biology Laboratory

High‐throughput automated DNA sequencing facility with fluorescent labels at the European Molecular Biology Laboratory

10.1002/elps.11501301125

Crossref journal-article

Wiley

ELECTROPHORESIS (311)

Abstract

AbstractOne of the aims of the facility is to develop and push the automated on‐line DNA sequencing gel technology to its limit in sequence throughput, which may be somewhere around 100 kilobases of sequence per device per day. Key new developments were initiated and applied in operation on the European Molecular Biology Laboratory (EMBL) automated sequencer and its commercial version A.L.F. (Pharmacia). Sequencing speed was increased by a factor of 10–20, up to 1500 bases per hour per clone on ultrathin (about 100 μm) gels, while the resolution and reading length were extended to 1000 bases on gels with 50 cm separation length, using fluorescein‐15‐*dATP as the internal label. With our sequencing strategy, closing about 40% of the sequence with “walking” primers and F‐15‐*dATP as internal label, we sequenced both strands of a cosmid insert of 38.5 kb in length, each strand twice, in only 430 sequencing reactions and with average reading of 380 bases per reaction.

Bibliography

Ansorge, W., Voss, H., Wiemann, S., Schwager, C., Sproat, B., Zimmermann, J., Stegemann, J., Erfle, H., Hewitt, N., & Rupp, T. (1992). Highâthroughput automated DNA sequencing facility with fluorescent labels at the European Molecular Biology Laboratory. ELECTROPHORESIS, 13(1), 616â619. Portico.

Authors 10

Wilhelm Ansorge (first)
Hartmut Voss (additional)
Stefan Wiemann (additional)
Christian Schwager (additional)
Brian Sproat (additional)
Jürgen Zimmermann (additional)
Josef Stegemann (additional)
Holger Erfle (additional)
Neil Hewitt (additional)
Thomas Rupp (additional)

References 14 Referenced 29

10.1016/S0021-9673(00)80076-X
10.1016/0003-2697(81)90031-2
10.1016/0165-022X(84)90064-2
10.1093/nar/19.3.675
{'key': 'e_1_2_1_6_2', 'first-page': '182', 'volume': '2', 'author': 'Stegemann J.', 'year': '1991', 'journal-title': 'Methods Mol. Cell. Biol.'} / Methods Mol. Cell. Biol. by Stegemann J. (1991)
10.1016/0165-022X(86)90038-2
{'key': 'e_1_2_1_8_2', 'first-page': '155', 'volume': '1', 'author': 'Voss H.', 'year': '1989', 'journal-title': 'Methods Mol. Cell. Biol.'} / Methods Mol. Cell. Biol. by Voss H. (1989)
10.1093/nar/18.6.1415
10.1093/nar/18.15.4417
10.1093/nar/15.11.4593
{'key': 'e_1_2_1_12_2', 'first-page': '30', 'volume': '3', 'author': 'Voss H.', 'year': '1992', 'journal-title': 'Methods Mol. Cell. Biol.'} / Methods Mol. Cell. Biol. by Voss H. (1992)
10.1093/nar/20.10.2471
Voss H. Wiemann S. Wirkner U. Schwager C. Zimmermann J. Stegemeann J. Erfle H. Hewitt N. A. Rupp T.andAnsorge W. Methods Mol. Cell. Biol.1992 in press.
10.1093/nar/18.4.1067

Dates

Type	When
Created	20 years, 3 months ago (May 28, 2005, 9:29 a.m.)
Deposited	1 year, 10 months ago (Oct. 24, 2023, 12:59 a.m.)
Indexed	1 year, 9 months ago (Nov. 17, 2023, 1:38 p.m.)
Issued	33 years, 8 months ago (Jan. 1, 1992)
Published	33 years, 8 months ago (Jan. 1, 1992)
Published Online	20 years, 4 months ago (April 13, 2005)
Published Print	33 years, 8 months ago (Jan. 1, 1992)

Funders 0

None

BibTeX

@article{Ansorge_1992, title={High‐throughput automated DNA sequencing facility with fluorescent labels at the European Molecular Biology Laboratory}, volume={13}, ISSN={1522-2683}, url={http://dx.doi.org/10.1002/elps.11501301125}, DOI={10.1002/elps.11501301125}, number={1}, journal={ELECTROPHORESIS}, publisher={Wiley}, author={Ansorge, Wilhelm and Voss, Hartmut and Wiemann, Stefan and Schwager, Christian and Sproat, Brian and Zimmermann, Jürgen and Stegemann, Josef and Erfle, Holger and Hewitt, Neil and Rupp, Thomas}, year={1992}, month=jan, pages={616–619} }

JSON

{
  "indexed": {
    "date-parts": [
      [
        2023,
        11,
        17
      ]
    ],
    "date-time": "2023-11-17T18:38:53Z",
    "timestamp": 1700246333082
  },
  "reference-count": 14,
  "publisher": "Wiley",
  "issue": "1",
  "license": [
    {
      "start": {
        "date-parts": [
          [
            2005,
            4,
            13
          ]
        ],
        "date-time": "2005-04-13T00:00:00Z",
        "timestamp": 1113350400000
      },
      "content-version": "vor",
      "delay-in-days": 4851,
      "URL": "http://onlinelibrary.wiley.com/termsAndConditions#vor"
    }
  ],
  "content-domain": {
    "domain": [],
    "crossmark-restriction": false
  },
  "published-print": {
    "date-parts": [
      [
        1992,
        1
      ]
    ]
  },
  "abstract": "<jats:title>Abstract</jats:title><jats:p>One of the aims of the facility is to develop and push the automated on\u2010line DNA sequencing gel technology to its limit in sequence throughput, which may be somewhere around 100 kilobases of sequence per device per day. Key new developments were initiated and applied in operation on the European Molecular Biology Laboratory (EMBL) automated sequencer and its commercial version A.L.F. (Pharmacia). Sequencing speed was increased by a factor of 10\u201320, up to 1500 bases per hour per clone on ultrathin (about 100 \u03bcm) gels, while the resolution and reading length were extended to 1000 bases on gels with 50 cm separation length, using fluorescein\u201015\u2010*dATP as the internal label. With our sequencing strategy, closing about 40% of the sequence with \u201cwalking\u201d primers and F\u201015\u2010*dATP as internal label, we sequenced both strands of a cosmid insert of 38.5 kb in length, each strand twice, in only 430 sequencing reactions and with average reading of 380 bases per reaction.</jats:p>",
  "DOI": "10.1002/elps.11501301125",
  "type": "journal-article",
  "created": {
    "date-parts": [
      [
        2005,
        5,
        28
      ]
    ],
    "date-time": "2005-05-28T13:29:23Z",
    "timestamp": 1117286963000
  },
  "page": "616-619",
  "source": "Crossref",
  "is-referenced-by-count": 29,
  "title": "High\u2010throughput automated DNA sequencing facility with fluorescent labels at the European Molecular Biology Laboratory",
  "prefix": "10.1002",
  "volume": "13",
  "author": [
    {
      "given": "Wilhelm",
      "family": "Ansorge",
      "sequence": "first",
      "affiliation": []
    },
    {
      "given": "Hartmut",
      "family": "Voss",
      "sequence": "additional",
      "affiliation": []
    },
    {
      "given": "Stefan",
      "family": "Wiemann",
      "sequence": "additional",
      "affiliation": []
    },
    {
      "given": "Christian",
      "family": "Schwager",
      "sequence": "additional",
      "affiliation": []
    },
    {
      "given": "Brian",
      "family": "Sproat",
      "sequence": "additional",
      "affiliation": []
    },
    {
      "given": "J\u00fcrgen",
      "family": "Zimmermann",
      "sequence": "additional",
      "affiliation": []
    },
    {
      "given": "Josef",
      "family": "Stegemann",
      "sequence": "additional",
      "affiliation": []
    },
    {
      "given": "Holger",
      "family": "Erfle",
      "sequence": "additional",
      "affiliation": []
    },
    {
      "given": "Neil",
      "family": "Hewitt",
      "sequence": "additional",
      "affiliation": []
    },
    {
      "given": "Thomas",
      "family": "Rupp",
      "sequence": "additional",
      "affiliation": []
    }
  ],
  "member": "311",
  "published-online": {
    "date-parts": [
      [
        2005,
        4,
        13
      ]
    ]
  },
  "reference": [
    {
      "key": "e_1_2_1_2_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1016/S0021-9673(00)80076-X"
    },
    {
      "key": "e_1_2_1_3_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1016/0003-2697(81)90031-2"
    },
    {
      "key": "e_1_2_1_4_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1016/0165-022X(84)90064-2"
    },
    {
      "key": "e_1_2_1_5_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1093/nar/19.3.675"
    },
    {
      "key": "e_1_2_1_6_2",
      "first-page": "182",
      "volume": "2",
      "author": "Stegemann J.",
      "year": "1991",
      "journal-title": "Methods Mol. Cell. Biol."
    },
    {
      "key": "e_1_2_1_7_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1016/0165-022X(86)90038-2"
    },
    {
      "key": "e_1_2_1_8_2",
      "first-page": "155",
      "volume": "1",
      "author": "Voss H.",
      "year": "1989",
      "journal-title": "Methods Mol. Cell. Biol."
    },
    {
      "key": "e_1_2_1_9_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1093/nar/18.6.1415"
    },
    {
      "key": "e_1_2_1_10_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1093/nar/18.15.4417"
    },
    {
      "key": "e_1_2_1_11_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1093/nar/15.11.4593"
    },
    {
      "key": "e_1_2_1_12_2",
      "first-page": "30",
      "volume": "3",
      "author": "Voss H.",
      "year": "1992",
      "journal-title": "Methods Mol. Cell. Biol."
    },
    {
      "key": "e_1_2_1_13_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1093/nar/20.10.2471"
    },
    {
      "key": "e_1_2_1_14_2",
      "unstructured": "Voss H. Wiemann S. Wirkner U. Schwager C. Zimmermann J. Stegemeann J. Erfle H. Hewitt N. A. Rupp T.andAnsorge W. Methods Mol. Cell. Biol.1992 in press."
    },
    {
      "key": "e_1_2_1_15_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1093/nar/18.4.1067"
    }
  ],
  "container-title": "ELECTROPHORESIS",
  "original-title": [],
  "language": "en",
  "link": [
    {
      "URL": "https://api.wiley.com/onlinelibrary/tdm/v1/articles/10.1002%2Felps.11501301125",
      "content-type": "unspecified",
      "content-version": "vor",
      "intended-application": "text-mining"
    },
    {
      "URL": "https://onlinelibrary.wiley.com/doi/full/10.1002/elps.11501301125",
      "content-type": "unspecified",
      "content-version": "vor",
      "intended-application": "similarity-checking"
    }
  ],
  "deposited": {
    "date-parts": [
      [
        2023,
        10,
        24
      ]
    ],
    "date-time": "2023-10-24T04:59:43Z",
    "timestamp": 1698123583000
  },
  "score": 1,
  "resource": {
    "primary": {
      "URL": "https://analyticalsciencejournals.onlinelibrary.wiley.com/doi/10.1002/elps.11501301125"
    }
  },
  "subtitle": [],
  "short-title": [],
  "issued": {
    "date-parts": [
      [
        1992,
        1
      ]
    ]
  },
  "references-count": 14,
  "journal-issue": {
    "issue": "1",
    "published-print": {
      "date-parts": [
        [
          1992,
          1
        ]
      ]
    }
  },
  "alternative-id": [
    "10.1002/elps.11501301125"
  ],
  "URL": "http://dx.doi.org/10.1002/elps.11501301125",
  "relation": {},
  "ISSN": [
    "0173-0835",
    "1522-2683"
  ],
  "subject": [],
  "container-title-short": "Electrophoresis",
  "published": {
    "date-parts": [
      [
        1992,
        1
      ]
    ]
  }
}