Miniprimer PCR, a New Lens for Viewing the Microbial World Export

@article{citeulike:2320213, abstract = {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 CR1 and CR2, suggesting that miniprimer PCR may reveal new dimensions of microbial diversity. 10.1128/AEM.01933-07}, author = {Isenbarger, Thomas A. and Finney, Michael and Rios-Velazquez, Carlos and Handelsman, Jo and Ruvkun, Gary}, citeulike-article-id = {2320213}, citeulike-linkout-0 = {http://dx.doi.org/10.1128/AEM.01933-07}, citeulike-linkout-1 = {http://aem.asm.org/content/74/3/840.abstract}, citeulike-linkout-2 = {http://aem.asm.org/content/74/3/840.full.pdf}, citeulike-linkout-3 = {http://aem.asm.org/cgi/content/abstract/74/3/840}, citeulike-linkout-4 = {http://view.ncbi.nlm.nih.gov/pubmed/18083877}, citeulike-linkout-5 = {http://www.hubmed.org/display.cgi?uids=18083877}, day = {1}, doi = {10.1128/AEM.01933-07}, issn = {1098-5336}, journal = {Appl. Environ. Microbiol.}, keywords = {16s, 16s\_rrna, pcr, thermo-rcdna, uncultured}, month = {February}, number = {3}, pages = {840--849}, posted-at = {2009-10-16 06:40:54}, priority = {2}, title = {Miniprimer PCR, a New Lens for Viewing the Microbial World}, url = {http://dx.doi.org/10.1128/AEM.01933-07}, volume = {74}, year = {2008} }

TY - JOUR ID - citeulike:2320213 L3 - citeulike-article-id:2320213 N2 - 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 CR1 and CR2, suggesting that miniprimer PCR may reveal new dimensions of microbial diversity. 10.1128/AEM.01933-07 IS - 3 JF - Appl. Environ. Microbiol. SN - 1098-5336 EP - 849 TI - Miniprimer PCR, a New Lens for Viewing the Microbial World VL - 74 SP - 840 KW - 16s KW - 16s_rrna KW - pcr KW - thermo-rcdna KW - uncultured AU - Isenbarger, Thomas AU - Finney, Michael AU - Rios-Velazquez, Carlos AU - Handelsman, Jo AU - Ruvkun, Gary PY - 2008/02/01/ UR - http://dx.doi.org/10.1128/AEM.01933-07 ER -