Sequins built for metagenomics.
Synthetic microbe genomes that together form a diverse community.
A synthetic microbial community.
Metagenomics can identify microbes within an environmental DNA sample, including microbes that are novel or uncultivable within the laboratory. However, the diversity of microbe sequences, the presence of host DNA, and errors and biases that accrue during the sequencing workflow can confound metagenome analysis.
We have built a set of sequins that represent a synthetic microbial community, and can be used as internal reference standards during metagenome sequencing and analysis.
Each sequin mirrors a sequence from a microbial genome. The sequins thereby retains the same nucleotide complexity, repetitiveness and sequencing performance of the original microbe genome.
Sequins represent a diverse range of microbe genomes from different taxa, with different genome sizes, GC content, rRNA operon count. Together, the sequin mixture represents a synthetic microbial community with phylogenetic and genomic heterogeneity.
Using sequins in metagenomics.
Sequins are ‘spiked-in’ to your extracted DNA sample. Together, the sample and sequins undergo library preparation and sequencing, where they accumulate the same errors and bias.
Following sequencing, the sequins can be distinguished from the accompanying sample in the output library by their synthetic sequence. The enables the sequins to be analyzed as internal quantitative and qualitative controls for each NGS library.
Sequins are mixed at staggered concentrations to form a reference ladder that can measure quantitative features in your NGS library. For example, sequins can determine the senstivity and accuracy for detecting microbes in a sample.
Errors and bias that accumulate during your metagenome experiment can also be measured with sequins. This allows benchmarking of lab protocols and bioinformatic methods, and more accurate normalization between samples.
These analysis (and more) can be performed using the anaquin software toolkit.
Synthetic microbe communities provide internal reference standards for metagenome sequencing and analysis. (2016)
Hardwick et. al.,
Using sequins with metagenome sequencing.