Journal
APPLIED AND ENVIRONMENTAL MICROBIOLOGY
Volume 74, Issue 3, Pages 840-849Publisher
AMER SOC MICROBIOLOGY
DOI: 10.1128/AEM.01933-07
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Funding
- NIAID NIH HHS [F32 AI065067, F32AI065067] Funding Source: Medline
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Molecular methods based on the 16S rRNA gene sequence are used widely in microbial ecology to reveal the diversity of microbial populations in environmental samples. Here we show that a new PCR method using an engineered polymerase and 10-nucleotide 'miniprimers expands the scope of detectable sequences beyond those detected by standard methods using longer primers and Taq polymerase. After testing the method in silico to identify divergent ribosomal genes in previously cloned environmental sequences, we applied the method to soil and microbial mat samples, which revealed novel 16S rRNA gene sequences that would not have been detected with standard primers. Deeply divergent sequences were discovered with high frequency and included representatives that define two new division-level taxa, designated CRI and CR2, suggesting that miniprimer PCR may reveal new dimensions of microbial diversity.
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