ARPHA Conference Abstracts : Conference Abstract

Print

Conference Abstract

Are multiplexed metabarcoding panels comparable to individual marker gene library preparations?

Niamh Eastwood^‡, Luisa Orsini^‡

‡ University of Birmingham, Birmingham, United Kingdom

Corresponding author: Niamh Eastwood (niamh.m.eastwood@gmail.com)

Received: 23 Feb 2021 | Published: 04 Mar 2021

© 2021 Niamh Eastwood, Luisa Orsini

This is an open access article distributed under the terms of the Creative Commons Attribution License (CC BY 4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

Citation: Eastwood N, Orsini L (2021) Are multiplexed metabarcoding panels comparable to individual marker gene library preparations? ARPHA Conference Abstracts 4: e64895. https://doi.org/10.3897/aca.4.e64895

Open Access

Abstract

Analysis of multiple marker genes using metabarcoding of environmental DNA (eDNA) can offer information greater than that from sequencing single marker genes, such as responses from across the phylogenetic tree to environmental gradients (Cordier et al. 2019). Furthermore, multiple regions of the same gene can be sequenced to improve phylogenetic resolution (Fuks et al. 2018). However, separate amplification reactions and library preparation steps for each marker can be costly and time consuming.

Here, we have designed and optimised a multiplex panel of four marker genes (two regions of 18S rRNA gene, one region of the 16S rRNA gene and one region of the rbcL gene). By combining steps into a single reaction, the labwork required is decreased, reducing cost and time. This multiplex is compared with a widely available commercial microbial (bacterial and fungal) screening panel and individual library preparations of each marker gene.

Keywords

Multiplex; Metabarcoding; eDNA;

Presenting author

Niamh Eastwood

Presented at

1st DNAQUA International Conference (March 9-11, 2021)

Acknowledgements

Funding program

Grant title

Hosting institution

University of Birmingham, UK

Ethics and security

Author contributions

Conflicts of interest

References

Cordier T, Frontalini F, Cermakova K, Apothloz-Perret-Gentil L, Treglia M, Scantamburlo E, Bonamin V, Pawlowski J (2019)

Multi-marker eDNA metabarcoding survey to assess the environmental impact of three offshore gas platforms in the North Adriatic Sea (Italy)

.

Marine Environmental Research

146

https://doi.org/10.1016/j.marenvres.2018.12.009

Fuks G, Elgart M, Amir A, Zeisel A, Turnbaugh P, Soen Y (2018)

Noam Shental Combining 16S rRNA gene variable regions enables high-resolution microbial community profiling

.

Microbiome

6

(

17

). https://doi.org/10.1186/s40168-017-0396-x

Supplementary material

Endnotes

Web
Analytics