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Updated: May 20, 2026

Pyrosequencing for Microbial Identification and Characterization
Published on: August 22, 2013
Errors in ribosomal sequence datasets generated using PCR-coupled 'panbacterial' pyrosequencing, and the
Erica M Prosdocimi1, Stefano Novati, Raffaele Bruno
1BTBS, Università degli Studi di Milano-Bicocca, Piazza della Scienza, 2, 20126 Milano, Italy.
Abstract:
Universal bacterial primers are often used in PCR-coupled sequencing approaches to investigate environmental and host-associated bacterial communities. Some of these primers can also amplify eukaryotic DNA. This is leading to the submission of datasets to public databases which are erroneously annotated as prokaryotic sequences. The present note sends a message about the risk of submitting incorrectly annotated sequence data and suggests a reliable approach for the sequencing of 16S rRNA genes and identification of bacteria within complex communities.
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