Amplicon Based 16S Ribosomal RNA Sequencing and Genus Identification
Poster Feb 19, 2015
*Jan Risinger, *Lindsey Renken, +Josh Hill, +Noushin Ghaffari, PhD, +Richard P. Metz, PhD, +Charles D. Johnson, PhD, *Masoud M. Toloue, Ph
Next generation sequencing analysis of 16S ribosomal RNA (rRNA) is commonly used to identify bacterial species and perform taxonomic studies. Bacterial 16S rRNA genes contain 9 hyper-variable regions with considerable sequence diversity among different bacterial species and can be used for species id. Rapid determination of highly complex bacterial populations through targeted amplification can provide an accurate gauge of diversity at taxonomic hierarchies as low as the genus level. A single 16S rRNA hypervariable domain does not have enough sequence diversity to distinguish genera. With increased read lengths of Illumina MiSeq chemistry, Bioo Scientific has expanded the common analysis of the fourth hyper-variable domain (V4) of prokaryotic 16S rRNA to V1, V2 and V3 regions simultaneously. Optimized preparation through a streamlined standardized procedure allows for high-quality, reproducible libraries. This optimization can be applied to different windows of 16S rRNA as well as other relevant prokaryotic taxonomic markers.
Despite the developments in conventional PCR, the complexity of multiplex Real Time PCR is still limited due to the lack of sufficient detection channels. To achieve high-end multiplexing capacity on standard Real Time PCR machines, Anapa Biotech has developed the MeltPlex® technology (see box on right).READ MORE