PCR-generated artefact from 16S rRNA gene-specific primers

Authors

  • Catherine A. Osborne,

    1. Department of Microbiology and Immunology, University of Melbourne, Parkville, Victoria 3010, Australia
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  • Maja Galic,

    1. Department of Microbiology and Immunology, University of Melbourne, Parkville, Victoria 3010, Australia
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  • Parveen Sangwan,

    1. Department of Microbiology and Immunology, University of Melbourne, Parkville, Victoria 3010, Australia
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  • Peter H. Janssen

    Corresponding author
    1. Department of Microbiology and Immunology, University of Melbourne, Parkville, Victoria 3010, Australia
      *Corresponding author. Tel.: +61 3 8344 5706; fax: +61 3 9347 1540., E-mail address: pjanssen@unimelb.edu.au
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  • Edited by C. Edwards

*Corresponding author. Tel.: +61 3 8344 5706; fax: +61 3 9347 1540., E-mail address: pjanssen@unimelb.edu.au

Abstract

Artefacts consisting of concatenated oligonucleotide primer sequences were generated during sub-optimally performing polymerase chain reaction amplification of bacterial 16S rRNA genes using a commonly employed primer pair. These artefacts were observed during amplification for terminal restriction fragment length polymorphism analyses of complex microbial communities, and after amplification from DNA from a microbial culture. Similar repetitive motifs were found in gene sequences deposited in GenBank. The artefact can be avoided by using different primers for the amplification reaction.

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