The Utilization of Linear Polylysine Coupled with Mechanic Forces to Extract Microbial DNA from Different Matrices

Celia François (1) , Celia Martinez (1) , Clement Faye (1) , Nathalie Pansu (2, 3) , Catherine Dunyach-Remy (2, 3) , Laurent Garrelly (1) , Benoit Roig (4) , Axelle Cadiere (4)

1 GL-biocontrol
2 CHU Nîmes - Centre Hospitalier Universitaire de Nîmes
3 VBMI - Virulence bactérienne et maladies infectieuses
4 CHROME - Détection, évaluation, gestion des risques CHROniques et éMErgents (CHROME) / Université de Nîmes

Molecular approaches are powerful tools that are used for medical or environmental diagnoses. However, the main limitations of such a tools are that they extract low levels of DNA and they do not remove the inhibitors of polymerase chain reaction (PCR). Although the use of polycation to complex and purify DNA has been described in the literature, elution often requires a high ionic strength or pH levels not compatible with molecular analyses. In this paper, we described a new process that is based on the complexation of DNA with linear polylysine, followed by capturing the complex by a cation exchange resin. The originality of the process consisted of using mechanic force to elute DNA from the complex. The extraction method showed several advantages when compared to existing methods, such as being compatible with pH levels that range from 5 to 11, as well as high levels of DNA recovery and elimination of PCR inhibitors from complex samples. This method was successfully applied to different types of samples, such as environmental samples, beverage samples, and medical samples. Furthermore, it was proven to be a good solution for removing PCR inhibitors and assuring good DNA recovery yield.

Mots clés: linear polylysine; purification DNA; PCR inhibitors; complex samples

Domaines: Biologie moléculaire

Celia François, Celia Martinez, Clement Faye, Nathalie Pansu, Catherine Dunyach-Remy, et al.. The Utilization of Linear Polylysine Coupled with Mechanic Forces to Extract Microbial DNA from Different Matrices. Microorganisms, 2020, 8 (12), pp.1901. ⟨10.3390/microorganisms8121901⟩. ⟨hal-03043809⟩