A direct PCR approach with low-biomass insert opens new horizons for molecular sciences on cryptogam communities

Author:

Jung Patrick1ORCID,Briegel-Williams Laura1,Werner Lina1,Jost Emily1,Schultz Matthias2,Nürnberg Dennis J.34,Grube Martin5,Lakatos Michael1

Affiliation:

1. Department of Integrative Biotechnology, University of Applied Sciences Kaiserslautern, Pirmasens, Germany

2. Institute for Plant Science and Microbiology, Herbarium Hamburgense, University of Hamburg, Hamburg, Germany

3. Institute of Experimental Physics, Freie Universität Berlin, Berlin, Germany

4. Dahlem Centre for Plant Sciences, Freie Universität Berlin, Berlin, Germany

5. Institute of Biology, University of Graz, Graz, Austria

Abstract

Cyanobacteria, green algae, lichens, and other cryptogams play crucial roles in complex microbial systems such as biological soil crusts of arid biomes or biofilms in caves. Molecular investigations on environmental samples or isolates of these microorganisms are often hampered by their dense aggregation, small size, or metabolism products which complicate DNA extraction and subsequent PCRs. Our work presents various examples of how a direct DNA extraction and PCR method relying on low biomass inserts can overcome these common problems and discusses additional applications of the workflow including adaptations.

Funder

Deutsche Forschungsgemeinschaft

Emmy Noether

Ministry of Science and Health RLP

Federal Ministry of Education and Research

Publisher

American Society for Microbiology

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