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Direct sequencing of long polymerase chain reaction fragments

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Abstract

Direct sequencing of polymerase chain reaction (PCR)-generated templates is a commonly used technique in molecular biology laboratories. We describe an improved method for direct sequencing of PCR fragments longer than 20 kb obtained with a commercial mixture ofTaq andPwo DNA polymerases. The sequencing protocol was optimized for an automated infrared DNA sequencer, consistently yielding long reads (500–600 bases).

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Authors and Affiliations

  1. Sezione di Microbiologia, Dipartimento di Biologia Molecolare, Università di Siena, Via Laterina 8, 53100, Siena, Italy

    Francesco Iannelli, Laura Giunti & Gianni Pozzi

Authors
  1. Francesco Iannelli
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  2. Laura Giunti
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  3. Gianni Pozzi
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Correspondence to Gianni Pozzi.

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Iannelli, F., Giunti, L. & Pozzi, G. Direct sequencing of long polymerase chain reaction fragments. Mol Biotechnol 10, 183–185 (1998). https://doi.org/10.1007/BF02760864

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  • DOI: https://doi.org/10.1007/BF02760864

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