Physical Isolation of Endospores from Environmental Samples by Targeted Lysis of Vegetative Cells
Tina Wunderlin, Thomas Junier, Christophe Paul, Nicole Jeanneret & Pilar Junier
| Résumé |
Endospore formation is a survival strategy found among some bacteria
from the phylum Firmicutes. During endospore formation, these
bacteria enter a morpho-physiological resting state that enhances
survival under adverse environmental conditions. Even though
endospore-forming Firmicutes are one of the most frequently
enriched and isolated bacterial groups in culturing studies, they
are often absent from diversity studies based on molecular methods.
The resistance of the spore core is considered one of the factors
limiting the recovery of DNA from endospores. We developed a method
that takes advantage of the higher resistance of endospores to
separate them from other cells in a complex microbial community
using physical, enzymatic and chemical lysis methods. The
endospore-only preparation thus obtained can be used for
re-culturing or to perform downstream analysis such as tailored DNA
extraction optimized for endospores and subsequent DNA sequencing.
This method, applied to sediment samples, has allowed the
enrichment of endospores and after sequencing, has revealed a large
diversity of endospore-formers in freshwater lake sediments. We
expect that the application of this method to other samples will
yield a similar outcome. |
| Citation | Wunderlin, T., Junier, T., Paul, C., Jeanneret, N., & Junier, P. (2016). Physical Isolation of Endospores from Environmental Samples by Targeted Lysis of Vegetative Cells. J Vis Exp(107). |
| Type | Article de périodique (Anglais) |
| Date de publication | 2016 |
| Nom du périodique | J Vis Exp |
| Numéro | 107 |