Development and Validation of Automated Methods for Metagenomic Analyses

Development and Validation of Automated Methods for Metagenomic Analyses

Russell Carmical, PhD, Assistant Professor, Baylor College of Medicine
Topic: Microbiology & Infectious Disease

Next Generation Sequencing (NGS) methods are arguably responsible for the establishment of the field of metagenomics. Significantly lower nucleic acid input requirements have made it possible to sample microbial communities that would have been deemed undetectable just a few years ago. In addition to technological advancements, the continued decline in the cost of sequencing has made it feasible to survey the thousands of samples necessary for metagenomic analyses. To meet the throughput demands of these studies, we have automated NGS sample processing methods from nucleic acid extraction, through library normalization, and pooling.

Here, we will present automated methods for 16S rDNA amplicon and whole genome shotgun sequencing as well as discuss the impact of automation on quality. The resulting sequencing reads from both approaches were compared for quality metrics using our standard quality matrix (Q30, Cluster Density, number of mapped reads, etc.)

Subsequently, sequencing reads were used for taxonomic identification and a comparative analysis. Although both automated methods offer a significant benefit in throughput, we found the reduction in the risk of human error far outweighs all other benefits. Furthermore, utilization of automated liquid handling reduces the risk of contamination.

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