Sequins built for metagenomics.
A diverse community of synthetic reference microbe genomes.
Design.
Metagenomics can identify microbes within an environmental DNA sample. However, the diversity of microbe sequences, the presence of host DNA, and technical errors and biases that accrue during the workflow can confound metagenome analysis.
We have built synthetic reference controls that represent a synthetic microbial community, and can be used as internal reference standards during metagenome sequencing and analysis.
Use.
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.
Analysis.
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.
Further reading.
Scientific paper
Synthetic microbe communities provide internal reference standards for metagenome sequencing and analysis. (2016) Hardwick et. al.,
Nature Communications.
Protocol
Using sequins with metagenome sequencing.
Laboratory protocol.
Version 5.2
March 2020
Review
Reference standards for next-generation sequencing.
(2017) Hardwick et. al.,
Nature Review Genetics.