Much of microbial taxonomy and metagenomic analyses nowadays are based on studies of the bacterial 16S ribosomal RNA gene (16S). 16S rRNA is a component of the 30S small subunit of the prokaryotic ribosome, which is an essential gene in all bacteria and archaea. Its corresponding gene is about 1500 bases in length, and it has both highly conserved and variable regions, which makes it an ideal taxonomic marker. Usually, primers that are designed to match conserved regions in the 16S gene are used to determine the complete DNA sequence via PCR amplification. The sequence of variable regions between the primers provides sufficient taxonomic information to match each amplified DNA fragment to a database of known 16S sequences and provides a putative identification for the organism that contained that DNA fragment in its genome. Most, if not all, bacteria can be differentiated by sequencing amplicons derived from one of the following variable regions: V1-V3, V3-V4 or V3-V5.
For study of metagenomics, 16S sequencing has great strength, since this method automatically selects only bacterial DNA for amplification and sequencing and it provides an extremely efficient taxonomically informative small marker sequence to measure the abundance of each taxon. Several tools have been developed for determining 16S metagenome sequences classification, such as SINA built into SILVA, Classifier in RDP and QIIME. The later QIIME wraps many packages which can be used to handle multiple processes and therefore has become the dominant tool.
16S rRNA Sequencing Services at Creative Biogene
Creative Biogene offers sequencing service for 16S rRNA gene from your bacterial or fungal colonies rapidly and effectively. To identify the microbial community in your sample, we provide one-stop service performing DNA extraction, PCR, sequencing and assembly to save your time at the bench.
Creative Biogene also provides services of advanced bioinformatic analysis, including i) 16S rRNA metagenome analysis; ii) Taxonomical assignment and read abundance estimation for all OTUs down to species level; iii) Abundance estimation of bacterial and archaeal OTUs considering lineage-specific copy numbers of marker genes. As an additional service, Creative Biogene will also isolate and purify the high molecular weight DNA from various starting material.
• 16S rRNA gene sequencing
• 16S rRNA metagenome analysis
• DNA isolation and purification
• Bacterial species identification
• High quality
• Fast and reliable turnaround
• Easily accessible, dedicated customer service
1. Hao, Y., Pei, Z., Brown, S.M. (2017) Bioinformatics in Microbiome Analysis. Methods in Microbiology. 44: 1-18.
Figure 1 Schematic representation of 16S rRNA of Escherichia coli.
(Methods in Microbiology 2017)