Science 2007,317(5846):1921–1926.PubMedCrossRef 33. Tumova P, Hofstetrova K, Nohynkova E, Hovorka O, Kral J: Cytogenetic evidence for diversity of two nuclei within a single diplomonad cell ofGiardia. Chromosoma 2007,116(1):65–78.PubMedCrossRef 34. Selmecki A, Forche A, Berman J: Aneuploidy and

isochromosome formation in drug-resistantCandida albicans. Science 2006,313(5785):367–370.PubMedCrossRef 35. Alby K, Bennett RJ: Sexual reproduction in theCandidaclade: cryptic cycles, diverse mechanisms, and alternative functions. Cell Mol Life Sci 2010,67(19):3275–3285.PubMedCrossRef Competing interests The authors declare that they have no competing interests. Author’s contribution JA and ML carried out the experiments and performed the data analyses. JA, ML and SGS contributed to the design and coordination of the experiments. JA wrote the manuscript. #learn more randurls[1|1|,|CHEM1|]# ML and SGS participated GDC-0994 cell line in editing the manuscript. All authors have read and approved the manuscript.”
“Background In the field of microbial ecology, the polymerase chain reaction (PCR) has been widely used for the amplification, detection and quantification of DNA targets since its introduction [1, 2], resulting in increased knowledge of the microbial world [3, 4]. However, the efficiency and accuracy of PCR can be diminished

by many factors including primer-template mismatches, reactant concentrations, the number of PCR cycles, annealing temperature, the complexity of the DNA template, and others. [5–7]. Primer-template mismatches are the most important because they can lead to selective amplification which prevents the correct assessment of microbial diversity

[8, 9]. Target sequences that cannot match the primers precisely will be amplified to a lesser extent, possibly even below the detection limit. The relative content of the sequences achieved is therefore changed, resulting in a deviation from the true community composition. Hence a comprehensive evaluation of bacterial primer coverage is critical to the interpretation of PCR results in microbial ecology research. Many related studies on primer coverage have been performed previously, but most are qualitative or semi-quantitative studies restricted to the domain MycoClean Mycoplasma Removal Kit level [10, 11]. Low coverage rates in some rare phyla might have been overlooked. Although Wang et al. [12] investigated primer coverage rates at the phylum level, only sequences from the Ribosomal Database Project (RDP) were used. This sole reliance on the RDP is another common limitation of previous studies. The RDP is a professional database containing more than one million 16S rRNA gene sequences. It also provides a series of data analysis services [13, 14], including Probe Match, which is often used in primer studies. However, despite the RDP’s large collection of sequences and extensive application, most of its sequences were generated through PCR amplification.