Next Generation Sequencing - Microbiome Deep Sequencing
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Deep Microbiome Sequencing
Next generation sequencing platforms (NGS) are currently revolutionizing the way researchers can study microbial colonization, infections and microbial communities (microbiomes). In microbiome research NGS has become a valuable tool for studying the microbial community [1], [2], [3]. Deep sequencing approaches have revealed new species and have given new insight into polymicrobial infections. Overall, deep sequencing of microbiomes has proven a huge potential for improving treatments (among others) against cystic fibrosis, gastric ulcer or vaginal infections. Over the past 25 years Sanger-based sequencing has been the “gold standard” and has been used for the characterization of microbiomes. Unfortunately Sanger sequencing is limited in terms of sequencing depth and sequence information is often limited to the most predominant species. In contrast NGS-based deep sequencing allows for in depth analysis of microbiomes on a genetic level. Nevertheless the depth of any microbiome deep sequencing approach is limited by factors like human DNA, reagents contaminations, and lack of calibrators. (Figure 1 summarizes the steps that can dramatically influence the quality of deep sequencing results, including sample processing, nucleic acid extraction and amplification). Furthermore it has not been possible so far to study sub-fractions of the microbiome (e.g. viable vs. dead microbiome; active vs. antibiotic-inhibited microbiome). Therefore, Molzym has now launched a range of innovative tools and services enabling the study of microbiomes.
Ultra-Deep Microbiome Sequencing - Unlocking the power of NGS
NGS-based microbiome bioanalysis aims to characterize the structure and dynamics of the living microbiome. DNA from dead microorganisms and human sources can significantly disturb the picture gained by deep sequencing results and significantly lowers the analytical sensitivity. Molzym offers a collection of research tools addressing current limitations in deep microbiome sequencing with respect to the removal of DNA from human cells and dead microbial cells, universal lysis of Gram-negative, Gram-positive and fungal organisms and supply of DNA-free extraction and PCR reagents.
Ultra-Deep Microbiome NGS Kit
The Ultra-Deep Microbiome NGS Kit is the ultimate solution combining protocols for extraction of enriched microbial DNA and amplification of bacterial and fungal sequences including positive controls, calibrators and DNA-free plastics. The Ultra-Deep Microbiome NGS Kit allows for significantly deeper sequencing compared to conventional DNA isolation and amplification.
Contamination-free PCR Reagents
Molzym’s enzyme purification technology removes traces of DNA from PCR reagents and supplies a highly active enzyme for research applications. This results in enhanced reliability in amplification of sequences originating from the community of bacteria and fungi. >> Go to contamination-free PCR reagents
Ultra-Deep Microbiome Service
Molzym runs a contract research lab developing NGS-assays and providing ultra-deep microbiome bioanalysis for the drug development industry as well as for academia. >> Go to the ultra-deep microbiome service
New White paper: Next Generation Sequencing: Ultra-Deep Microbiome Sequencing
This new white paper gives an overview on factors limiting the analytical sensitivity of NGS-based microbiome approaches and introduces innovative tools enabling ultra-deep microbiome bioanalysis. >>Download PDF
References
[1] The Human Microbiome Project: http://commonfund.nih.gov/hmp/index.aspx
[2] Siqueira JF et al. (2012) J. Oral Microbiol. 4:10743-???
[3] Rogers GB, Bruce KD (2010) Mol. Diagn. Ther. 14:343-350