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Transposon-facilitated DNA sequencing

Strathmann, Michael and Hamilton, Bruce A. and Mayeda, Carol A. and Simon, Melvin I. and Meyerowitz, Elliot M. and Palazollo, Michael J. (1991) Transposon-facilitated DNA sequencing. Proceedings of the National Academy of Sciences of the United States of America, 88 (4). pp. 1247-1250. ISSN 0027-8424. PMCID PMC50994. doi:10.1073/pnas.88.4.1247.

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We describe here a transposon-based DNA sequencing strategy that allows the introduction of sequencing priming sites throughout a target sequence by bacterial mating. A miniplasmid was designed to select against transposon insertions into the vector. Sites of transposon insertion are mapped by the polymerase chain reaction with bacterial overnight cultures providing the templates. A small set of plasmids with transposons spaced several hundred base pairs apart can then be sequenced. Sequencing primers corresponding to the transposon ends allow sequencing in both directions. Thus, the entire sequence of both strands can be easily determined.

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Meyerowitz, Elliot M.0000-0003-4798-5153
Additional Information:© 1991 National Academy of Sciences Communicated by E. B. Lewis, September 26, 1990. We thank K. VijayRaghavan, Susan Celniker, and Christopher Martin for many helpful discussions. We also thank Douglas Berg for ATnSeq5 and Randy Reed for the sequence of GD1 and GD2. M.J.P. is a Lucille P. Markey Scholar of the Lucille P. Markey Charitable Trust. B.A.H. is supported, in part, by a U.S. Public Health Service Predoctoral National Research Service Award (T32 GM07616). This work was also supported by U.S. Public Health Service Program Project Grant GM40499 (E.M.M.) and by U.S. Department of Energy Grant FG03-89ER60891 (M.I.S.). The publication costs of this article were defrayed in part by page charge payment. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. §1734 solely to indicate this fact.
Funding AgencyGrant Number
Lucille P. Markey Charitable TrustUNSPECIFIED
NIH Predoctoral FellowshipT32 GM07616
Department of Energy (DOE)DE-FG03-89ER60891
Subject Keywords:[gamma][delta], polymerase chain reaction
Issue or Number:4
PubMed Central ID:PMC50994
Record Number:CaltechAUTHORS:STRpnas91
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Official Citation:M Strathmann, B A Hamilton, C A Mayeda, M I Simon, E M Meyerowitz, and M J Palazzolo Transposon-facilitated DNA sequencing. PNAS 1991 88 (4) 1247-1250; doi:10.1073/pnas.88.4.1247
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:661
Deposited By: Archive Administrator
Deposited On:12 Sep 2005
Last Modified:08 Nov 2021 19:04

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