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.
The novel ‘K’ DNA extraction protocol offers a reliable alternative for preparing fecal specimens for 16S rRNA gene amplicon sequencing that maintains representation of microbial populations in a sample, with the added benefits that the K method reduces hands-on time by up to 20 x for 96 sample preparations.READ MORE