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

2005; Oxford University Press; Volume: 248; Issue: 2 Linguagem: Inglês

10.1016/j.femsle.2005.05.043

1574-6968

Catherine A. Osborne, Maja Galić, Parveen Sangwan, Peter H. Janssen,

Microbial Community Ecology and Physiology

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.